Veterinary Sciences and One Health Journal VSOHJ (original) (raw)

Papers by Veterinary Sciences and One Health Journal VSOHJ

Veterinary Sciences and One Health Journal, 2024

Canine infestation of ectoparasites is of medical and veterinary importance worldwide and in Iran... more Canine infestation of ectoparasites is of medical and veterinary importance worldwide and in Iran due to transmission of pathogen agents. The present study was aimed to determine prevalence and species diversity of flea in dogs of Tabriz city, Iran. A total of 100 stray dogs were randomly selected and examined from March 2021 to March 2022. Kind of fleas were collected from body surface of examined dogs and identified. The results indicated that 24 percent (24%) of stray dog were infested with ectoparasites. There was a significant difference between prevalence and sex in dogs. The maximum and minimum fleas' infestations were respectively for Ctenocephalides canis (14%) and Xenopsylla cheopis (2%). Dogs under study infected with dog fleas (14%) out of all examined dogs were infested with Ctenocephalides canis, Ctenocephalides felis (5%), pulex irritans (3%), Xenopsylla cheopis (2%). The prevalence of fleas in stray dogs in Tabriz is high, and the necessary health care should be taken seriously to control and prevent them, because the possibility of contaminating the environment and humans by dog fleas is high.

Veterinary Sciences and One Health Journal, 2024

Although anti-parasitic drugs in humans and animals have been used for many years to control para... more Although anti-parasitic drugs in humans and animals have been used for many years to control parasitic diseases in developed, developing and lowincome countries. However, the continued use of anti-parasitic drugs is associated with two basic problems, which include the gradual resistance of the parasite to the drug and its residue in animal products such as meat and milk, and the pollution of the environment.Therefore, according to the mentioned points, researchers tried to use vaccines to solve the problems caused by drug consumption, although conventional vaccines and recombinant vaccines have been associated with effective results in preventing the spread of bacterial and viral diseases in humans and animals, and in the case of vaccines Recombination related to parasitic diseases brilliant research activities have been carried out so far. But no significant results have been obtained. Despite this, a limited number of protozoan vaccines such as Theilerian, Babesian, Coccidial, Toxoplasmic and Leishmanian vaccines have been prepared and produced in the form of attenuated vaccines, but the use of such vaccines is accompanied by problems and obstacles.In the case of molecular parasitic vaccines, vaccines with high reproductive immunity have not yet been prepared and investigations are ongoing in this regard, and in the current situation, only anti-mite vaccines such as Tick Gard in Australia and Gavac in Cuba and others. It is used in South American countries, but it should be noted that the success rate of such vaccines is not more than 30-35%, and it is necessary to conduct more studies to prepare anti-mite molecular vaccines with an efficiency of more than 35%.

Veterinary Sciences and One Health Journal, 2024

Bovine babesiosis is the disease which is caused by hemolytic protozoan babesia species (particul... more Bovine babesiosis is the disease which is caused by hemolytic protozoan babesia species (particularly B. bovis and B. bigemina). It causes the huge economic loss in the livestock industry of the world. Currently, control measures are insufficient to prevent the animals from this disease. Hence, the aim of this study is to construct the multi-epitope antigenic fragment of B. bigemina and to express its mRNA which can be used as a potent vaccine material in future against bovine babesiosis, as mRNA is more potent and economical than other vaccines. It will go for immediate translation of the antigen. Various potential antigens of Babesia bigemina (AMA-1, P58, GP45, Rap-1α) from already reported vaccine candidate proteins were analyzed through NCBI database tool (https://www.ncbi.nlm.nih.gov/). Then, the consensus sequences of these proteins were generated by using software; DNASTAR and epitope prediction was done by using various immuno-informatic tools like VaxiJen server, immune epitope database, NetCTL server v1.2 and ABCpred tool for prediction of antigenicity, CTL epitope, HTL epitope and B-cell epitope prediction analysis. Multi-epitope construct was formed based on synthetic DNA. Then, cloning of that multi-epitope fragment was done into T7 promoter containing plasmid through transfection into DH5α competent cells. Plasmid was extracted from the liquid phase of lysate through the elution-absorption process on the matrix of silica. Restriction analysis as well as linearization of the plasmid was done. By using Megascript in-vitro transcription kit, mRNA from the linearized plasmid was transcribed. Quantification of mRNA was done by Regression analysis. After production of mRNA, it was purified by using mRNA purification kit (43). Multi-epitope fragment of B. bigemina was constructed and it can be used to transcribe in-vitro. Resulting mRNA could replace the other vaccines as it can immediately translate into antigenic protein without incorporating host genome and will provide more promising results than other conventional vaccines.

Veterinary Sciences and ne Health Journal, 2024

Parasites of genus Lernaea are responsible for a disease named lernaeosis in freshwater fish host... more Parasites of genus Lernaea are responsible for a disease named lernaeosis in freshwater fish hosts. Identification of Lernaea based on the shape of its anchors is ill-defined. Moreover, morphological plasticity of Lernaea has made its identification very difficult. In this study, 25 Labeo rohita samples were examined for Lernaea. Three Lernaea parasites were recovered and were identified based on the morphology of their anchors. They were identified to be Lernaea cruciate. Partial 18S rRNA gene sequence of parasites was determined. BLAST results showed that the parasites were 94.15% similar to other Lernaea sequences present in GenBank database. Phylogenetic tree generated by neighbor-joining method showed that our Lernaea clustered separately within the same clade with other Lernaea isolates from GenBank. Both BLAST results and phylogenetic analysis suggest that Lernaea from our study is a different species than other Lernaea species present in GenBank but to reach a solid conclusion more molecular work involving different genes and hosts are needed. It is suggested to sequence of more genes of Lernaea from vast range of hosts in order to resolve taxonomy of this genus. This is the first molecular study of Lernaea from Pakistan.

Veterinary Sciences and One Health Journal, 2024

Bovine theileriosis is a devastating disease of livestock, i.e. cattle, which causes huge economi... more Bovine theileriosis is a devastating disease of livestock, i.e. cattle, which causes huge economic losses in terms of mortality, cost of treatment and productivity, all over the world including Pakistan. It is transmitted by biological tick vectors of the genus Hyalomma. Out of the various methods to control bovine theileriosis, vaccination is ranked to be the most effective method. A total of 20 infected blood samples were collected from cattle and buffaloes, 10 from each. Similarly, 20 samples of lymph nodes were collected from cattle and buffaloes. Detection of theileriosis in lymph nodes was done using PCR. After that Lymphocytes were separated out through density gradient centrifugation in case of blood samples and with the help crushing and filtration with nylon cloth in case of lymph nodes. Lymphocytes were counted through hemocytometer. Before culture, lymphocytes were counted per ml of the culture medium and then by using RPMI-1640 culture medium these lymphocytes from all the 40 samples (including both the blood and lymph node samples from each of the cattle and buffaloes) were cultured by using MASP technique for Theileria infected bovine lymphocytes culture. Initially, there were many factors which either compromised or disturbed the culture viabilities. The factors included were fungal and microbial contamination, most probably because of frequent interventions of technical persons and others in the cell culture laboratory to install some equipment and or to solve the problems arisen because of malfunctioning of some laboratory equipment and has been considered as the most likely cause of contamination seen in the lymphocyte culture during the current study. To determine if there was the difference between groups, cell count of 40 cell lines at different time points, repeated measures analysis of variance (ANOVA) was applied by considering time as a factor. It was revealed that there was at least one group whose mean was different from another group (p-value <0.0005). To investigate, which group means were different from other groups, a post hoc multiple comparisons was applied. It was revealed that all combinations were statistically different from each other (p-value < 0.005). We have applied repeated measures analysis of variance (ANOVA) by taking time as a factor. The results revealed that there was at least one group whose mean was different from another group (p-value < 0.005, df=4, F statistic 250.05). To determine which group was different from other group, a post-hoc multiple comparison using Scheffe method was applied. This analysis revealed that all combinations were statistically different (p-value 0.005).

Veterinary Sciences and One Health Journal, 2024

Equines are very important animals because of their role in defense, transport and sports. The eq... more Equines are very important animals because of their role in defense, transport and sports. The equines are neglected as compared to other food animals in spite of their role in supporting poor people especially in Pakistan. In recent years, many equines have been observed suffering from a chronic debilitating disease that somehow has resemblance to some extent to chronic parasitic diseases like Leishmaniosis and Trypanosomiasis but the animals don't respond to any parasitic treatment. Amastigote like bodies were demonstrated in the blood of these animals in microscopy, hence the current study investigated the nature of these amastigote like bodies using PCR. The blood samples of the equines carried to the Department of Parasitology of University of Veterinary and Animal Sciences Lahore, from the different public organizations in Lahore, were used for this study. The samples having amastigote like bodies verified through microscopy were subjected to DNA based diagnosis. DNA extracted with commercially available kit or conventional method was subjected to PCR by using genus specific primers that target the protozoan parasites i.e. Leishmania and Babesia. 38 samples were found having amastigote like bodies in the smears, out of a total of 97. Most of the animals fell ill in winter season as compared to other seasons. 25 samples out of the ones positive in microscopy were subjected to PCR targeting Leishmania major (ITS gene), Leishmania tropica (kinetoplastid gene) and Leishmania donovani (small subunit rDNA gene) but no amplification was observed. These 25 samples were then subjected to PCR targeting Babesia bigemina (18sRNA) where amplification was seen only in two samples. These two samples were got sequenced; out of that only one sequence fell in alignment with the sequence of Babesia bigemina from other parts of the world particularly from Spain, indicating the possible nature of amastigote like bodies as to be arisen from Babesia bigemina, a protozoan parasite causing babesiosis in bovines. Presence of Babesia bigemina in equines raises many questions about the host specificity of Babesia and related parasites. Before drawing any solid conclusions about the host specificity, further investigations are warranted by targeting more genes of Babesia bigemina, followed by a detailed sequence analysis of all targeted genes.

Veterinary Sciences and One Health Journal, 2024

Trypanosomiasis is a protozoan disease that is transmitted by biting flies. Trypanosomes species,... more Trypanosomiasis is a protozoan disease that is transmitted by biting flies. Trypanosomes species, including Trypanosome evansi (T. evansi) and Trypanosome theileri (T. theileri) have been reported in a wide range of mammalian species worldwide. Owing to the paucity of information, the current study was designed to investigate the prevalence and genomic characterization of T. evansi and T. theileri in the cattle population in Southern Punjab, Pakistan. Therefore, a robust surveillance system is needed to monitor this disease for the accurate diagnosis and understand the prevalence of each species. Here, we explained the detection of trypanosome species through deep amplicon sequencing in co-infection communities and made the concept of haemoprotobiome. The DNA artificial pools from four different Trypanosoma isolates (T. theileri, T. vivax, T. congolense, and T. brucei) were created, and the threshold of the deep amplicon sequencing method and the proportion of each of the species present or absent in the mixer, 3 replicates of 10 "artificial pools mixture" was evaluated. It was revealed that this method could determine the accurate composition of Trypanosoma species in the pooled samples. Second, the technique was applied to validate the detection threshold of Trypanosoma infected samples collected from cattle and were identified as an infection of Trypanosoma using PCR-based assay. Each sample shows the different composition of Trypanosoma species, and the amount of Illumina Mi-Seq reads in the samples where the mixed infection was present. Third, we applied the assay to the field samples to new insight into the species composition of Trypanosoma communities in cattle, camel, buffalo, horse, sheep, and goat in the endemic region of Pakistan. It was confirmed that T. evansi is the primary species in Pakistan, and T. theileri has been observed to reveal the first report of this group of parasites in this country. In this study, we used molecular genetic approaches to provide relevant insight into the emergence of diminazene resistance after intensive positive selection pressure at the AT-1 locus of T. evansi in the endemic regions of Pakistan. We chose this region because animals are treated sporadically often with generic diminazene drugs of unknown quality. Diminazene drugs have been widely used for the therapy of Trypanosomiasis because they are relatively safe and inexpensive. Until recently, it was assumed that diminazene was effective against T. evansi, but the widespread resistance shows the need to investigate the emergence of resistance in this group of parasites. There is a clear need to understand how the resistance mutations against diminazene drugs emerge. The frequency with which diminazene resistance in T. evansi emerges is important for its prevention and control Our findings show that this technology can be used in disease surveillance programs in animals and humans, similar to how our approach revolutionized the research of piroplasms in humans and bovines. These works explore the practical methods which are helpful to determine the epidemiology of different diseases, including Trypanosoma and co-infection. The technology is useful for monitoring the parasite diversity after the emergence and the drug resistance in cattle. The plasticity of this approach is not host-specific and could have future application for the assessment of other animals and human Trypanosoma species.

Veterinary Sciences and One Health Journal, 2023

Trypanosomiasis is the most significant disease of camels across the globe. It causes huge econom... more Trypanosomiasis is the most significant disease of camels across the globe. It causes huge economic as well as animal health losses in Pakistan. Trypanosomiasis is commonly caused by Trypanosoma evansi; however other Trypanosoma species can also produce the disease but have not been reported in Pakistan, yet. The outbreaks and prevalence of the disease is still not documented in Baluchistan province. Trypanosoma evansi is thought to be responsible for such high mortality, which resulted in great economic and production losses. In the current study 100 camel samples (27 male and 73 female) from three tehsils of Lasbela district were examined using microscopic and molecular (PCR) examination. For microscopic study 0.2 ml blood was collected from ear vein and for molecular examination 5 ml blood was collected from jugular vein. The PCR based prevalence was found to be 20%. Two cases were recognized microscopically and 5 were confirmed by PCR examination. PCR has much higher diagnostic sensitivity than the conventional microscopic examination. It improves the species specificity in diagnosis of trypanosomiasis compared to microscopic examination. The infection was more prevalent in the adult camels than the younger ones and the infection was more prevalent in Lasi breed compared to Makrani breed. This study confirms that trypanosomiasis is a possible threat, affecting the health and productivity of camels, in particular adult animals. To study the husbandry practices of camel herds a questionnaire was designed which included data on farm type, camel population age and sex of the animals, educational status of the farmer, flies control strategy, housing material, feeding practices are included in the study. The data described that all the farmers (n=12) were uneducated, most of the farmers had dependency for their livelihoods on livestock and agriculture business. More than 70% of farmers kept local breed. All of them practiced pasture based farming system. They said that flies burden were high in summer season and they neither applied any flies control measure nor any veterinary team ever visited their farm.

Veterinary Sciences and One Health Journal, 2023

After parturition, endometritis is the most prevalent and important disease of dairy animals. Und... more After parturition, endometritis is the most prevalent and important disease of dairy animals. Understanding of common uterine pathogens and risk factors are necessary for the improvement of reproductive health at herd or country level. In the last few decades excessive and inappropriate use and dosage of intrauterine medicines resulted in development of drug resistance against the bacteria. The drug residues of some antibiotics in milk have a lot of importance in dairy industry. The instant study was planned to find out the antibacteriall & therapeutic efficacy of pomegranate, garlic & turmeric alone in relation to most effective antibiotic selected on the basis of antibiogram against the isolates from cervical mucus obtained from repeat breeding crossbred cows suffering from endometritis. In-vitro antibacterial activity of the different herbal extracts were determined by Disc diffusion method against Staphylococcus aureus in cervical mucus of repeat breeding crossbred cows. The minimum inhibitory concentration (MIC) of these herbal extracts was also ascertained by micro dilution method. A total of 32 endometritic cows were selected on the basis of history, per-rectal examination, and white side test of cervical mucus. Each sample primarily was cultured onto the Nutrient agar. Then select media was used for the separation of Staphylococcus aureus. Then AST of different antibiotics and herbal extracts (pomegranate, garlic, and turmeric) were checked against the Staphylococcus aureus. Then five groups were made (A, B, C, D&E), each group allotted five animals. Group A received a 100ml pomegranate. Group B received 100ml turmeric. Group C received 100ml garlic. Group D received 40ml gentamicin. Group E act as control group was given 100ml N.S. All the extracts, antibiotic and normal saline were injected via intra-uterine route. Cervical mucus samples of repeater cows were collected aseptically on the day of estrus before treatment and tested for bacterial load. Blood samples were taken on the day of estrus before treatment and at post treatment subsequent estrus for hematological studies (Hb, PCV, RBCs, Neutrophils, lymphocytes & monocytes) to know the health status of animals. In-vitro antibacterial activities of herbal extracts were tested and compared with antibacterial activity of Gentamicin. Result showed that the ethanolic extract of pomegranate showed antibacterial against Staphylococcus aureus, whereas aqueous extract of garlic and ethanolic extract of turmeric showed no antibacterial activity against Staphylococcus aureus.

Veterinary Sciences and One Health, 2023

The seroprevalence of toxoplasmosis ranges widely in various areas of the world and different pop... more The seroprevalence of toxoplasmosis ranges widely in various areas of the world and different populations. However, toxoplasmosis is commonly benign and asymptomatic; it causes major signs and symptoms of toxoplasmosis in immune compromised individuals and especially in pregnant women during pregnancy. Prevention of congenital infection depends upon the awareness of association of factors to cause T. gondii infection. The preventive measures depend on the women's knowledge about toxoplasmosis. The purpose of the study was to check the knowledge on association factors to cause toxoplasmosis among pet owners in Punjab, Pakistan. The cross-sectional study was done, multicenter 2019 and June 2022 in Pet Center, University of Veterinary and Animal Sciences (UVAS), Lahore and other different cities of Punjab, Pakistan. Overall, 238 pet owners were asked to participate in the survey of toxoplasmosis. Of those, 202 agreed, which resulted in the participation rate of 84.87%. The potential association of factors to induce the toxoplasmosis was determined using multivariate logistic regression analysis. According to questionnaire, have you any cat in your house (yes), place of residence (OR, 4.94; 95% CI, 1.55-1.56; p< 0.000),who handles the boxes of cats, advanced age (OR, 1.73; 95% CI, 1.73-2.92;p < 0.041), have you any abortion history in your family (yes), place of residence (OR, 4.18; 95% CI, 2.15-8.14; p< 0.000), have you any house member having schizophrenia (yes), place of residence (OR, 5.36; 95% CI, 1.22-2.35; p< 0.000) turned out significantly associated with better to all others questions of questionnaire of toxoplasmosis. Among pet owners in Punjab, Pakistan, the strong association of cat among pet owners (OR, 4.94) and abortion in pregnant women was found due to relation with cats (OR, 4.18) to cause toxoplasmosis. Younger age, city residence and higher education level brought out significantly associated with better knowledge of association factors for T. gondii infection.

Veterinary Sciences and One Health Journal, 2023

Livestock plays a vital role in socioeconomic development of Pakistan through poverty alleviation... more Livestock plays a vital role in socioeconomic development of Pakistan through poverty alleviation. Livestock contributed 60.66% in agri-business and 11% in GDP. The expansion of livestock sector's net worth increases from 1430 billion to 1466 billion in last two years. Bovine (49.6 million cattle and 41.2 million buffalo) are the two main milk and meat producing animals with approximately milk (59 billion tons) and beef production (23 million tons), respectively. Anaplasmosis cause heavy economic losses in livestock sector including high morbidity, production and reproduction losses. The study will be conducted on a commercial dairy farm of various age group located at the suburbs of Sargodha region. The sample size (n=130) cattle dairy breed was considered for screening of parasites through fecal and blood microscopy. The screened animals were divided into two groups having parasitic infection and those free from parasitic infestation. The focused parameters were Parasitemia, milk production, feeding, and type of feeding, feed cost, treatment cost and labor cost, respectively. Anaplasma causes significant economic loss of both dairy and beef industry in livestock. With yearly loss of 64millionindairycattle(64 million in dairy cattle (64millionindairycattle(60 per cow) estimated in Costa Rica by anaplasmosis. In Latin America, estimated economic loss is $875 million per annum due to anaplasmosis. Cattle become carriers of A. marginale after their initial infection and act as a reservoir to keep the bacterium alive in their herds. Our study concluded that 22.92% economic losses on a commercial dairy farm due to blood parasites. Anaplasma was highly prevalent among parasitic infections. The parasitic infection impacted the farm profitability negatively with medicine cost and decrease in milk production had the highest impact. Anaplasma was highly prevalent among parasitic infections. The parasitic infection impacted the farm profitability negatively with medicine cost and decrease in milk production had the highest impact. Farm-level economic impact was evaluated which provides baseline information about the disease losses which could be used to assess the economic efficiency of parasitic control interventions. The information will help dairy farmers to modify their agricultural techniques as well as policymakers and decision makers to control anaplasmosis at farm level in Punjab in specific and country in general.

Veterinary Sciences and One Health Journal, 2023

To date, there have been no reported cases of Anaplasma marginale infection found in both the blo... more To date, there have been no reported cases of Anaplasma marginale infection found in both the blood and ticks of the buffalo. A 7.5-year-old female buffalo showed progressive weakness, anemia, high fever (104°F), tachycardia, labored breathing, pale mucous membranes, and tick infestation. Further investigation through microscopy, hematology, and PCR confirmed that it was suffering from A. marginale. A thin blood smear showed dot forms of A. marginale at the margins of stained RBCs. The parasitemia was found to be 16%. Blood hematological analysis revealed decreased RBC count, HCT, PCV and MCHC, PLT, WBC count, Lymphocyte count, and monocyte count. The ticks found on the buffalo were identified as Hyalomma anatolicum anatolicum. PCR tests on both the blood and tick samples positive for A. marginale. The buffalo was treated with oxytetracycline at a dose of 30ml/250kg body weight slow I/M daily for five days and Amivit at a dose of 20ml IM for three consecutive days. Clinical signs started to subside three days post-treatment, and complete recovery from parasitemia and symptoms was achieved by seven weeks post-treatment.

Veterinary Sciences and One Health Journal, 2023

Ticks belong to arachnida which is class of phylum arthropoda. Ticks have two major families Ixod... more Ticks belong to arachnida which is class of phylum arthropoda. Ticks have two major families Ixodidae (hard ticks) and Argasidae (soft ticks). In livestock mostly hard ticks transmit various diseases including Anaplasmosis, Babesiosis and Theileriosis. Theileriosis is disease caused by Theileria that is apicomplexan parasite and is transmitted by Hyalomma tick. Theileria annulata is protozoan parasite that causes tropical theileriosis in cattle. It causes morbidity and mortality in cattle which results in considerable economic losses. To reduce these losses control of disease is very necessary. Vector (ticks) control or vaccine can help in control of disease. There are various types of vaccines Killed vaccine and live vaccine, live vaccine provides high protection against homologous strains but very low protection against heterologous strain. There are issues related with cold chain maintenance in live vaccine. There is need of effective vaccine against Theileriosis. Recombinant vaccine is one of them. This study was initiative for development of vaccine. T. annulata was detected through PCR and isolated from salivary glands of Hyalomma ticks. The methodology employed the use of isolated sporozoites of T. annulata in a cattle calf vaccination trial. Adult female Hyalomma ticks were gathered from a specific geographic location. These ticks underwent careful cleaning and dissection to isolate their salivary glands, which were preserved for analysis. After pooling the glands, the combined sample was utilized to detect T. annulata. DNA extraction and PCR amplification were conducted using specific techniques and primers to identify the presence of the target gene. The sporozoites of T. annulata were then isolated using a specialized protocol and further examined under a microscope to confirm their presence through staining and molecular analysis.

Veterinary Sciences and One Health Journal, 2023

Malaria is a parasitic disease spread by Anopheles mosquitos and caused by Plasmodium protozoan s... more Malaria is a parasitic disease spread by Anopheles mosquitos and caused by Plasmodium protozoan species. Plasmodium falciparum, widely known as the "king of illnesses," is the parasite that causes the most deadly and lethal form of malaria. There are also many human malaria species, such as P. knowlesi, P. vivax, P. ovale, and P. malariae that are responsible for acute disease, severe sickness, but they have a very low fatality rate. In subtropical and tropical areas, malaria is the most common infectious disease. Malaria remains a major public health concern, impacting over 500 million people each year and resulting in 1-2 million deaths, 90% of whom are children from Sub-Saharan Africa. To help with diagnosis, image analysis software and artificial intelligence-based machine learning approaches were used to measure parasitemia in microscopic blood slides. The current study compared the efficacy of an automated malaria screener to a polymerase chain reaction for the detection of Plasmodium falciparum to examine the performance of this novel mobile application in detecting Plasmodium falciparum. 50 blood samples from suspected malaria patients were collected in EDTA tube from different hospitals in Lahore. The collected samples were screened for detection of Plasmodium falciparum by using Malaria screener and polymerase chain reaction (PCR). PCR method was considered as the reference method for the diagnosis of malaria. Out of 50 samples, 30 (60%) were identified as positive (P. vivax) by the analysis of blood smears by malaria screener and only 18 (36%) were identified positive for Plasmodium vivax and none was found positive for Plasmodium falciparum. The sensitivity and specificity of malaria screener were calculated and found to be 50% and 77.77%, respectively. Cohen's kappa coefficient was used to compare agreement between malaria screener and PCR, considered as reference standard. When comparing the malaria screener and the PCR, this study discovered that there was moderate agreement (k=0.49).

Veterinary Sciences and One Health Journal, 2023

Brucellosis is one of the most important zoonotic diseases worldwide. Brucellosis is not only haz... more Brucellosis is one of the most important zoonotic diseases worldwide. Brucellosis is not only hazardous to human but also causes a huge economic loss due to abortions in cattle, buffaloes, sheep and goats. Three main species are involved in brucellosis, i.e. Brucella melitensis in goats and sheep, Brucella suis in pigs, and in cattle Brucella abortus. The present study was designed to serologically diagnose and to find the associated risk factors of brucellosis in Punjab, Pakistan. In a total of buffaloes 100 and human 20 serum samples were randomly collected from different dairy farms, and were screened for brucellosis by Rose-Bengal Plate Test (RBPT). The positive RBPT samples were examined by indirect ELISA. Data of all animals was recorded on predesigned data captured form, which included age, sex, breed, housing system, feeding system, management, BCS, vaccination etc. Overall occurrence of Brucella abortus was recorded as 15 %(15/100) by RBPT and 5% by indirect ELISA in buffaloes. Moreover, in humans it was 5% (1/20) by RBPT and 0% by indirect ELISA. The higher occurrence was recorded in more than 5 years old age buffaloes, with larger herd sizes having an abortion history and retained placenta. This is the first study exploring buffaloes and human brucellosis in Punjab. The outcomes of this study will be helpful for farmers and veterinarians in evolving effective control plans.

Veterinary Sciences and One Health Journal, 2023

The study was conducted to evaluate efficacy of controlled internal drug releasing device (CIDR) ... more The study was conducted to evaluate efficacy of controlled internal drug releasing device (CIDR) and to assess levels of hormone i.e. Progesterone and minerals i.e. Calcium, Phosphorus during synchronization in postpartum anestrous Nili-Ravi buffaloes during low breeding season. For this purpose, a total of twenty buffaloes were divided into two groups each with 10 animal in each group. For the treatment group, CIDR was inserted intra-vaginal for 14 days and an injection of 2ml Dalmazine (150 µg d-cloprostenol) per animal was administered by intramuscularly on day 13. After CIDR removal, animals were observed for estrus and subsequently inseminated 12 and 24 hours after standing heat. Pregnancy was assessed at day 60 post insemination. In both groups, blood sampling was done at day 0, 7, 9, 11, 14, and 21 Post Artificial Insemination (AI) for analysis of Progesterone, calcium and phosphorus and difference was calculated between treatment and control group. Further, their difference was evaluated between pregnant and non-pregnant buffaloes. The progesterone concentration was measured through ELISA while levels of calcium and phosphorus were measured through spectrophotometry. The estrus intensity, ovulation rate and pregnancy rate were significant (P<0.05) higher in group A buffaloes. It was concluded that CIDR can be used efficiently for induction of estrus during low breeding season in buffaloes.

Veterinary Sciences and One Health Journal, 2023

In livestock, chronic diarrhoea is one of the clinical disorders associated with copper deficienc... more In livestock, chronic diarrhoea is one of the clinical disorders associated with copper deficiency. The purpose of this study will be to survey copper deficiency related chronic diarrhea in cattle and buffaloes in urban and peri-urban areas of Faisalabad. Study animals (n = 384) will be selected by unknown status of prevalence, the sample size was calculated by keeping the 50% expected prevalence with 95% confidence limit. A detailed proforma was designed and used to investigate the type of fodders and mineral supplements given to the animals in the study area. Blood samples were collected from animals suffering from chronic diarrhea in the study area. Sera samples were used to assess copper levels by atomic absorption spectrophotometry at Hi Tech Laboratory, University of Agriculture Faisalabad. Prevalence of chronic diarrhoea in cattle and buffaloes in the study area was calculated by simple binomial method and Chi square test were used to determine copper levels between diarrhoeic and non-diarrhoeic animals.

Veterinary Sciences and One Health Journal, 2023

N. caninum is an animal parasitic protozoan. Neospora can persist in the majority of dairy cows i... more N. caninum is an animal parasitic protozoan. Neospora can persist in the majority of dairy cows is facilitated by its vertical transmission. N. caninuminduced abortion in bovines is a significant contributor to economic losses in Pakistan's burgeoning dairy sector. Research comprises four brain samples of aborted bovine fetuses from several regions in the province of Punjab. The objective of this study was the isolation and propagation of the parasite, which is the first step toward the production of a vaccine against this parasite. Microscopic and molecular identification of N. caninum was performed through PCR. N. caninum was isolated from the brains of aborted fetal samples, propagated in the mice and then mice were killed to take brains, Neospora parasite was confirmed using PCR. Continuation of the experimentation meant for isolation and propagation by exploiting dams to be found seropositive in the planned studies, will be an important perspective of the study.

Veterinary Sciences and One Health Journal, 2023

Haemonchosis is responsible for a wide range of health problems, economic losses in small ruminan... more Haemonchosis is responsible for a wide range of health problems, economic losses in small ruminants and is characterized by anemia, dehydration, loss of appetite and body condition, retarded growth, and reduced production performance. Haemonchus Contortus has develops benzimidazole resistance. Total of 100 samples (n = 50 sheep, n = 50 goats) was collected directly from animals' abomasum. Collection of samples was done from the local abattoir of Shahpur Kanjra Lahore (Pakistan). The genomic DNA of adult male H. contortus was extracted by using DNA extraction kit and phenol chloroform method. Pair of primers was developed for H. contortus. After that, the PCR product was run on 1.2% agarose gel. The product was compared with known marker of DNA ladder to have correct amplified band. The PCR amplified band was purify by using gel extraction kit and sent with primers for sequencing. The sequences were aligned and analyzed by BIOEDIT software. By this alignment tool, mutations in different codon were observed and compared with β tubulin gene sequence from gene bank of NCBI. Significance of mutations at different locations was evaluated by using Chi Square Test. Data on mutation in H. contortus was analyzed by Pearson's chisquare test by using SPSS version 17. The results obtained showing 0.004% significance in sheep. While no mutation was observed in goats. That is no significant as compared to sheep samples. Percentages of negative and positive samples were calculated. Mutations in different codon in β tubulin gene were observed and their percentages were compared. By finding the mutations, this study helped to trace out the BZ's resistance in H. contortus in local sheep and goats and also determine the changes at codon 200 of sheep.

Veterinary Sciences and One Health Journal, 2023

Wild and domestic bovids are susceptible for obligate blood parasites consisting of Theileria, Ba... more Wild and domestic bovids are susceptible for obligate blood parasites consisting of Theileria, Babesia and Anaplasma. These are the most economically important and lethal parasites in dairy industry. The haemoparasites prevalence depends on managemental factors such as locality, herd size, tick control strategies, tick infestation rate and housing system which have been neglected. Therefore, the current study was designed to investigate the correlation of managemental practices with prevalence of haemoparasites. For this purpose, a total of 50 (30 before and 20 after medication) blood samples were collected from cattle of physically and clinically showing the symptoms of haemoparasites infection. Upon microscopic examination, the prevalence of theileriosis, babesiosis, anaplasmosis and mix infection was 40% (9/30), 30% (7/30) and 23.33% (5/30) and 20% (6/30) respectively before medication. After medication with buparvaquone and imidocarb dipropionate combined with oxytetracycline, the blood samples of those animals were again processed for microscopic examination. The prevalence of theileriosis, babesiosis and anaplasmosis was 30% (6/20), 20% (4/20) and 10% (2/20), respectively after 24-48 hour of treatment. The medication was effective to treat the infected animals but lacks managemental practices were the source of re-infection. It is concluded that only treatment of haemoparasites is not the way to control disease but necessary to improve the management eradicate the habitat of ticks.

Veterinary Sciences and One Health Journal, 2024

Canine infestation of ectoparasites is of medical and veterinary importance worldwide and in Iran... more Canine infestation of ectoparasites is of medical and veterinary importance worldwide and in Iran due to transmission of pathogen agents. The present study was aimed to determine prevalence and species diversity of flea in dogs of Tabriz city, Iran. A total of 100 stray dogs were randomly selected and examined from March 2021 to March 2022. Kind of fleas were collected from body surface of examined dogs and identified. The results indicated that 24 percent (24%) of stray dog were infested with ectoparasites. There was a significant difference between prevalence and sex in dogs. The maximum and minimum fleas' infestations were respectively for Ctenocephalides canis (14%) and Xenopsylla cheopis (2%). Dogs under study infected with dog fleas (14%) out of all examined dogs were infested with Ctenocephalides canis, Ctenocephalides felis (5%), pulex irritans (3%), Xenopsylla cheopis (2%). The prevalence of fleas in stray dogs in Tabriz is high, and the necessary health care should be taken seriously to control and prevent them, because the possibility of contaminating the environment and humans by dog fleas is high.

Veterinary Sciences and One Health Journal, 2024

Although anti-parasitic drugs in humans and animals have been used for many years to control para... more Although anti-parasitic drugs in humans and animals have been used for many years to control parasitic diseases in developed, developing and lowincome countries. However, the continued use of anti-parasitic drugs is associated with two basic problems, which include the gradual resistance of the parasite to the drug and its residue in animal products such as meat and milk, and the pollution of the environment.Therefore, according to the mentioned points, researchers tried to use vaccines to solve the problems caused by drug consumption, although conventional vaccines and recombinant vaccines have been associated with effective results in preventing the spread of bacterial and viral diseases in humans and animals, and in the case of vaccines Recombination related to parasitic diseases brilliant research activities have been carried out so far. But no significant results have been obtained. Despite this, a limited number of protozoan vaccines such as Theilerian, Babesian, Coccidial, Toxoplasmic and Leishmanian vaccines have been prepared and produced in the form of attenuated vaccines, but the use of such vaccines is accompanied by problems and obstacles.In the case of molecular parasitic vaccines, vaccines with high reproductive immunity have not yet been prepared and investigations are ongoing in this regard, and in the current situation, only anti-mite vaccines such as Tick Gard in Australia and Gavac in Cuba and others. It is used in South American countries, but it should be noted that the success rate of such vaccines is not more than 30-35%, and it is necessary to conduct more studies to prepare anti-mite molecular vaccines with an efficiency of more than 35%.

Veterinary Sciences and One Health Journal, 2024

Bovine babesiosis is the disease which is caused by hemolytic protozoan babesia species (particul... more Bovine babesiosis is the disease which is caused by hemolytic protozoan babesia species (particularly B. bovis and B. bigemina). It causes the huge economic loss in the livestock industry of the world. Currently, control measures are insufficient to prevent the animals from this disease. Hence, the aim of this study is to construct the multi-epitope antigenic fragment of B. bigemina and to express its mRNA which can be used as a potent vaccine material in future against bovine babesiosis, as mRNA is more potent and economical than other vaccines. It will go for immediate translation of the antigen. Various potential antigens of Babesia bigemina (AMA-1, P58, GP45, Rap-1α) from already reported vaccine candidate proteins were analyzed through NCBI database tool (https://www.ncbi.nlm.nih.gov/). Then, the consensus sequences of these proteins were generated by using software; DNASTAR and epitope prediction was done by using various immuno-informatic tools like VaxiJen server, immune epitope database, NetCTL server v1.2 and ABCpred tool for prediction of antigenicity, CTL epitope, HTL epitope and B-cell epitope prediction analysis. Multi-epitope construct was formed based on synthetic DNA. Then, cloning of that multi-epitope fragment was done into T7 promoter containing plasmid through transfection into DH5α competent cells. Plasmid was extracted from the liquid phase of lysate through the elution-absorption process on the matrix of silica. Restriction analysis as well as linearization of the plasmid was done. By using Megascript in-vitro transcription kit, mRNA from the linearized plasmid was transcribed. Quantification of mRNA was done by Regression analysis. After production of mRNA, it was purified by using mRNA purification kit (43). Multi-epitope fragment of B. bigemina was constructed and it can be used to transcribe in-vitro. Resulting mRNA could replace the other vaccines as it can immediately translate into antigenic protein without incorporating host genome and will provide more promising results than other conventional vaccines.

Veterinary Sciences and ne Health Journal, 2024

Parasites of genus Lernaea are responsible for a disease named lernaeosis in freshwater fish host... more Parasites of genus Lernaea are responsible for a disease named lernaeosis in freshwater fish hosts. Identification of Lernaea based on the shape of its anchors is ill-defined. Moreover, morphological plasticity of Lernaea has made its identification very difficult. In this study, 25 Labeo rohita samples were examined for Lernaea. Three Lernaea parasites were recovered and were identified based on the morphology of their anchors. They were identified to be Lernaea cruciate. Partial 18S rRNA gene sequence of parasites was determined. BLAST results showed that the parasites were 94.15% similar to other Lernaea sequences present in GenBank database. Phylogenetic tree generated by neighbor-joining method showed that our Lernaea clustered separately within the same clade with other Lernaea isolates from GenBank. Both BLAST results and phylogenetic analysis suggest that Lernaea from our study is a different species than other Lernaea species present in GenBank but to reach a solid conclusion more molecular work involving different genes and hosts are needed. It is suggested to sequence of more genes of Lernaea from vast range of hosts in order to resolve taxonomy of this genus. This is the first molecular study of Lernaea from Pakistan.

Veterinary Sciences and One Health Journal, 2024

Bovine theileriosis is a devastating disease of livestock, i.e. cattle, which causes huge economi... more Bovine theileriosis is a devastating disease of livestock, i.e. cattle, which causes huge economic losses in terms of mortality, cost of treatment and productivity, all over the world including Pakistan. It is transmitted by biological tick vectors of the genus Hyalomma. Out of the various methods to control bovine theileriosis, vaccination is ranked to be the most effective method. A total of 20 infected blood samples were collected from cattle and buffaloes, 10 from each. Similarly, 20 samples of lymph nodes were collected from cattle and buffaloes. Detection of theileriosis in lymph nodes was done using PCR. After that Lymphocytes were separated out through density gradient centrifugation in case of blood samples and with the help crushing and filtration with nylon cloth in case of lymph nodes. Lymphocytes were counted through hemocytometer. Before culture, lymphocytes were counted per ml of the culture medium and then by using RPMI-1640 culture medium these lymphocytes from all the 40 samples (including both the blood and lymph node samples from each of the cattle and buffaloes) were cultured by using MASP technique for Theileria infected bovine lymphocytes culture. Initially, there were many factors which either compromised or disturbed the culture viabilities. The factors included were fungal and microbial contamination, most probably because of frequent interventions of technical persons and others in the cell culture laboratory to install some equipment and or to solve the problems arisen because of malfunctioning of some laboratory equipment and has been considered as the most likely cause of contamination seen in the lymphocyte culture during the current study. To determine if there was the difference between groups, cell count of 40 cell lines at different time points, repeated measures analysis of variance (ANOVA) was applied by considering time as a factor. It was revealed that there was at least one group whose mean was different from another group (p-value <0.0005). To investigate, which group means were different from other groups, a post hoc multiple comparisons was applied. It was revealed that all combinations were statistically different from each other (p-value < 0.005). We have applied repeated measures analysis of variance (ANOVA) by taking time as a factor. The results revealed that there was at least one group whose mean was different from another group (p-value < 0.005, df=4, F statistic 250.05). To determine which group was different from other group, a post-hoc multiple comparison using Scheffe method was applied. This analysis revealed that all combinations were statistically different (p-value 0.005).

Veterinary Sciences and One Health Journal, 2024

Equines are very important animals because of their role in defense, transport and sports. The eq... more Equines are very important animals because of their role in defense, transport and sports. The equines are neglected as compared to other food animals in spite of their role in supporting poor people especially in Pakistan. In recent years, many equines have been observed suffering from a chronic debilitating disease that somehow has resemblance to some extent to chronic parasitic diseases like Leishmaniosis and Trypanosomiasis but the animals don't respond to any parasitic treatment. Amastigote like bodies were demonstrated in the blood of these animals in microscopy, hence the current study investigated the nature of these amastigote like bodies using PCR. The blood samples of the equines carried to the Department of Parasitology of University of Veterinary and Animal Sciences Lahore, from the different public organizations in Lahore, were used for this study. The samples having amastigote like bodies verified through microscopy were subjected to DNA based diagnosis. DNA extracted with commercially available kit or conventional method was subjected to PCR by using genus specific primers that target the protozoan parasites i.e. Leishmania and Babesia. 38 samples were found having amastigote like bodies in the smears, out of a total of 97. Most of the animals fell ill in winter season as compared to other seasons. 25 samples out of the ones positive in microscopy were subjected to PCR targeting Leishmania major (ITS gene), Leishmania tropica (kinetoplastid gene) and Leishmania donovani (small subunit rDNA gene) but no amplification was observed. These 25 samples were then subjected to PCR targeting Babesia bigemina (18sRNA) where amplification was seen only in two samples. These two samples were got sequenced; out of that only one sequence fell in alignment with the sequence of Babesia bigemina from other parts of the world particularly from Spain, indicating the possible nature of amastigote like bodies as to be arisen from Babesia bigemina, a protozoan parasite causing babesiosis in bovines. Presence of Babesia bigemina in equines raises many questions about the host specificity of Babesia and related parasites. Before drawing any solid conclusions about the host specificity, further investigations are warranted by targeting more genes of Babesia bigemina, followed by a detailed sequence analysis of all targeted genes.

Veterinary Sciences and One Health Journal, 2024

Trypanosomiasis is a protozoan disease that is transmitted by biting flies. Trypanosomes species,... more Trypanosomiasis is a protozoan disease that is transmitted by biting flies. Trypanosomes species, including Trypanosome evansi (T. evansi) and Trypanosome theileri (T. theileri) have been reported in a wide range of mammalian species worldwide. Owing to the paucity of information, the current study was designed to investigate the prevalence and genomic characterization of T. evansi and T. theileri in the cattle population in Southern Punjab, Pakistan. Therefore, a robust surveillance system is needed to monitor this disease for the accurate diagnosis and understand the prevalence of each species. Here, we explained the detection of trypanosome species through deep amplicon sequencing in co-infection communities and made the concept of haemoprotobiome. The DNA artificial pools from four different Trypanosoma isolates (T. theileri, T. vivax, T. congolense, and T. brucei) were created, and the threshold of the deep amplicon sequencing method and the proportion of each of the species present or absent in the mixer, 3 replicates of 10 "artificial pools mixture" was evaluated. It was revealed that this method could determine the accurate composition of Trypanosoma species in the pooled samples. Second, the technique was applied to validate the detection threshold of Trypanosoma infected samples collected from cattle and were identified as an infection of Trypanosoma using PCR-based assay. Each sample shows the different composition of Trypanosoma species, and the amount of Illumina Mi-Seq reads in the samples where the mixed infection was present. Third, we applied the assay to the field samples to new insight into the species composition of Trypanosoma communities in cattle, camel, buffalo, horse, sheep, and goat in the endemic region of Pakistan. It was confirmed that T. evansi is the primary species in Pakistan, and T. theileri has been observed to reveal the first report of this group of parasites in this country. In this study, we used molecular genetic approaches to provide relevant insight into the emergence of diminazene resistance after intensive positive selection pressure at the AT-1 locus of T. evansi in the endemic regions of Pakistan. We chose this region because animals are treated sporadically often with generic diminazene drugs of unknown quality. Diminazene drugs have been widely used for the therapy of Trypanosomiasis because they are relatively safe and inexpensive. Until recently, it was assumed that diminazene was effective against T. evansi, but the widespread resistance shows the need to investigate the emergence of resistance in this group of parasites. There is a clear need to understand how the resistance mutations against diminazene drugs emerge. The frequency with which diminazene resistance in T. evansi emerges is important for its prevention and control Our findings show that this technology can be used in disease surveillance programs in animals and humans, similar to how our approach revolutionized the research of piroplasms in humans and bovines. These works explore the practical methods which are helpful to determine the epidemiology of different diseases, including Trypanosoma and co-infection. The technology is useful for monitoring the parasite diversity after the emergence and the drug resistance in cattle. The plasticity of this approach is not host-specific and could have future application for the assessment of other animals and human Trypanosoma species.

Veterinary Sciences and One Health Journal, 2023

Trypanosomiasis is the most significant disease of camels across the globe. It causes huge econom... more Trypanosomiasis is the most significant disease of camels across the globe. It causes huge economic as well as animal health losses in Pakistan. Trypanosomiasis is commonly caused by Trypanosoma evansi; however other Trypanosoma species can also produce the disease but have not been reported in Pakistan, yet. The outbreaks and prevalence of the disease is still not documented in Baluchistan province. Trypanosoma evansi is thought to be responsible for such high mortality, which resulted in great economic and production losses. In the current study 100 camel samples (27 male and 73 female) from three tehsils of Lasbela district were examined using microscopic and molecular (PCR) examination. For microscopic study 0.2 ml blood was collected from ear vein and for molecular examination 5 ml blood was collected from jugular vein. The PCR based prevalence was found to be 20%. Two cases were recognized microscopically and 5 were confirmed by PCR examination. PCR has much higher diagnostic sensitivity than the conventional microscopic examination. It improves the species specificity in diagnosis of trypanosomiasis compared to microscopic examination. The infection was more prevalent in the adult camels than the younger ones and the infection was more prevalent in Lasi breed compared to Makrani breed. This study confirms that trypanosomiasis is a possible threat, affecting the health and productivity of camels, in particular adult animals. To study the husbandry practices of camel herds a questionnaire was designed which included data on farm type, camel population age and sex of the animals, educational status of the farmer, flies control strategy, housing material, feeding practices are included in the study. The data described that all the farmers (n=12) were uneducated, most of the farmers had dependency for their livelihoods on livestock and agriculture business. More than 70% of farmers kept local breed. All of them practiced pasture based farming system. They said that flies burden were high in summer season and they neither applied any flies control measure nor any veterinary team ever visited their farm.

Veterinary Sciences and One Health Journal, 2023

After parturition, endometritis is the most prevalent and important disease of dairy animals. Und... more After parturition, endometritis is the most prevalent and important disease of dairy animals. Understanding of common uterine pathogens and risk factors are necessary for the improvement of reproductive health at herd or country level. In the last few decades excessive and inappropriate use and dosage of intrauterine medicines resulted in development of drug resistance against the bacteria. The drug residues of some antibiotics in milk have a lot of importance in dairy industry. The instant study was planned to find out the antibacteriall & therapeutic efficacy of pomegranate, garlic & turmeric alone in relation to most effective antibiotic selected on the basis of antibiogram against the isolates from cervical mucus obtained from repeat breeding crossbred cows suffering from endometritis. In-vitro antibacterial activity of the different herbal extracts were determined by Disc diffusion method against Staphylococcus aureus in cervical mucus of repeat breeding crossbred cows. The minimum inhibitory concentration (MIC) of these herbal extracts was also ascertained by micro dilution method. A total of 32 endometritic cows were selected on the basis of history, per-rectal examination, and white side test of cervical mucus. Each sample primarily was cultured onto the Nutrient agar. Then select media was used for the separation of Staphylococcus aureus. Then AST of different antibiotics and herbal extracts (pomegranate, garlic, and turmeric) were checked against the Staphylococcus aureus. Then five groups were made (A, B, C, D&E), each group allotted five animals. Group A received a 100ml pomegranate. Group B received 100ml turmeric. Group C received 100ml garlic. Group D received 40ml gentamicin. Group E act as control group was given 100ml N.S. All the extracts, antibiotic and normal saline were injected via intra-uterine route. Cervical mucus samples of repeater cows were collected aseptically on the day of estrus before treatment and tested for bacterial load. Blood samples were taken on the day of estrus before treatment and at post treatment subsequent estrus for hematological studies (Hb, PCV, RBCs, Neutrophils, lymphocytes & monocytes) to know the health status of animals. In-vitro antibacterial activities of herbal extracts were tested and compared with antibacterial activity of Gentamicin. Result showed that the ethanolic extract of pomegranate showed antibacterial against Staphylococcus aureus, whereas aqueous extract of garlic and ethanolic extract of turmeric showed no antibacterial activity against Staphylococcus aureus.

Veterinary Sciences and One Health, 2023

The seroprevalence of toxoplasmosis ranges widely in various areas of the world and different pop... more The seroprevalence of toxoplasmosis ranges widely in various areas of the world and different populations. However, toxoplasmosis is commonly benign and asymptomatic; it causes major signs and symptoms of toxoplasmosis in immune compromised individuals and especially in pregnant women during pregnancy. Prevention of congenital infection depends upon the awareness of association of factors to cause T. gondii infection. The preventive measures depend on the women's knowledge about toxoplasmosis. The purpose of the study was to check the knowledge on association factors to cause toxoplasmosis among pet owners in Punjab, Pakistan. The cross-sectional study was done, multicenter 2019 and June 2022 in Pet Center, University of Veterinary and Animal Sciences (UVAS), Lahore and other different cities of Punjab, Pakistan. Overall, 238 pet owners were asked to participate in the survey of toxoplasmosis. Of those, 202 agreed, which resulted in the participation rate of 84.87%. The potential association of factors to induce the toxoplasmosis was determined using multivariate logistic regression analysis. According to questionnaire, have you any cat in your house (yes), place of residence (OR, 4.94; 95% CI, 1.55-1.56; p< 0.000),who handles the boxes of cats, advanced age (OR, 1.73; 95% CI, 1.73-2.92;p < 0.041), have you any abortion history in your family (yes), place of residence (OR, 4.18; 95% CI, 2.15-8.14; p< 0.000), have you any house member having schizophrenia (yes), place of residence (OR, 5.36; 95% CI, 1.22-2.35; p< 0.000) turned out significantly associated with better to all others questions of questionnaire of toxoplasmosis. Among pet owners in Punjab, Pakistan, the strong association of cat among pet owners (OR, 4.94) and abortion in pregnant women was found due to relation with cats (OR, 4.18) to cause toxoplasmosis. Younger age, city residence and higher education level brought out significantly associated with better knowledge of association factors for T. gondii infection.

Veterinary Sciences and One Health Journal, 2023

Livestock plays a vital role in socioeconomic development of Pakistan through poverty alleviation... more Livestock plays a vital role in socioeconomic development of Pakistan through poverty alleviation. Livestock contributed 60.66% in agri-business and 11% in GDP. The expansion of livestock sector's net worth increases from 1430 billion to 1466 billion in last two years. Bovine (49.6 million cattle and 41.2 million buffalo) are the two main milk and meat producing animals with approximately milk (59 billion tons) and beef production (23 million tons), respectively. Anaplasmosis cause heavy economic losses in livestock sector including high morbidity, production and reproduction losses. The study will be conducted on a commercial dairy farm of various age group located at the suburbs of Sargodha region. The sample size (n=130) cattle dairy breed was considered for screening of parasites through fecal and blood microscopy. The screened animals were divided into two groups having parasitic infection and those free from parasitic infestation. The focused parameters were Parasitemia, milk production, feeding, and type of feeding, feed cost, treatment cost and labor cost, respectively. Anaplasma causes significant economic loss of both dairy and beef industry in livestock. With yearly loss of 64millionindairycattle(64 million in dairy cattle (64millionindairycattle(60 per cow) estimated in Costa Rica by anaplasmosis. In Latin America, estimated economic loss is $875 million per annum due to anaplasmosis. Cattle become carriers of A. marginale after their initial infection and act as a reservoir to keep the bacterium alive in their herds. Our study concluded that 22.92% economic losses on a commercial dairy farm due to blood parasites. Anaplasma was highly prevalent among parasitic infections. The parasitic infection impacted the farm profitability negatively with medicine cost and decrease in milk production had the highest impact. Anaplasma was highly prevalent among parasitic infections. The parasitic infection impacted the farm profitability negatively with medicine cost and decrease in milk production had the highest impact. Farm-level economic impact was evaluated which provides baseline information about the disease losses which could be used to assess the economic efficiency of parasitic control interventions. The information will help dairy farmers to modify their agricultural techniques as well as policymakers and decision makers to control anaplasmosis at farm level in Punjab in specific and country in general.

Veterinary Sciences and One Health Journal, 2023

To date, there have been no reported cases of Anaplasma marginale infection found in both the blo... more To date, there have been no reported cases of Anaplasma marginale infection found in both the blood and ticks of the buffalo. A 7.5-year-old female buffalo showed progressive weakness, anemia, high fever (104°F), tachycardia, labored breathing, pale mucous membranes, and tick infestation. Further investigation through microscopy, hematology, and PCR confirmed that it was suffering from A. marginale. A thin blood smear showed dot forms of A. marginale at the margins of stained RBCs. The parasitemia was found to be 16%. Blood hematological analysis revealed decreased RBC count, HCT, PCV and MCHC, PLT, WBC count, Lymphocyte count, and monocyte count. The ticks found on the buffalo were identified as Hyalomma anatolicum anatolicum. PCR tests on both the blood and tick samples positive for A. marginale. The buffalo was treated with oxytetracycline at a dose of 30ml/250kg body weight slow I/M daily for five days and Amivit at a dose of 20ml IM for three consecutive days. Clinical signs started to subside three days post-treatment, and complete recovery from parasitemia and symptoms was achieved by seven weeks post-treatment.

Veterinary Sciences and One Health Journal, 2023

Ticks belong to arachnida which is class of phylum arthropoda. Ticks have two major families Ixod... more Ticks belong to arachnida which is class of phylum arthropoda. Ticks have two major families Ixodidae (hard ticks) and Argasidae (soft ticks). In livestock mostly hard ticks transmit various diseases including Anaplasmosis, Babesiosis and Theileriosis. Theileriosis is disease caused by Theileria that is apicomplexan parasite and is transmitted by Hyalomma tick. Theileria annulata is protozoan parasite that causes tropical theileriosis in cattle. It causes morbidity and mortality in cattle which results in considerable economic losses. To reduce these losses control of disease is very necessary. Vector (ticks) control or vaccine can help in control of disease. There are various types of vaccines Killed vaccine and live vaccine, live vaccine provides high protection against homologous strains but very low protection against heterologous strain. There are issues related with cold chain maintenance in live vaccine. There is need of effective vaccine against Theileriosis. Recombinant vaccine is one of them. This study was initiative for development of vaccine. T. annulata was detected through PCR and isolated from salivary glands of Hyalomma ticks. The methodology employed the use of isolated sporozoites of T. annulata in a cattle calf vaccination trial. Adult female Hyalomma ticks were gathered from a specific geographic location. These ticks underwent careful cleaning and dissection to isolate their salivary glands, which were preserved for analysis. After pooling the glands, the combined sample was utilized to detect T. annulata. DNA extraction and PCR amplification were conducted using specific techniques and primers to identify the presence of the target gene. The sporozoites of T. annulata were then isolated using a specialized protocol and further examined under a microscope to confirm their presence through staining and molecular analysis.

Veterinary Sciences and One Health Journal, 2023

Malaria is a parasitic disease spread by Anopheles mosquitos and caused by Plasmodium protozoan s... more Malaria is a parasitic disease spread by Anopheles mosquitos and caused by Plasmodium protozoan species. Plasmodium falciparum, widely known as the "king of illnesses," is the parasite that causes the most deadly and lethal form of malaria. There are also many human malaria species, such as P. knowlesi, P. vivax, P. ovale, and P. malariae that are responsible for acute disease, severe sickness, but they have a very low fatality rate. In subtropical and tropical areas, malaria is the most common infectious disease. Malaria remains a major public health concern, impacting over 500 million people each year and resulting in 1-2 million deaths, 90% of whom are children from Sub-Saharan Africa. To help with diagnosis, image analysis software and artificial intelligence-based machine learning approaches were used to measure parasitemia in microscopic blood slides. The current study compared the efficacy of an automated malaria screener to a polymerase chain reaction for the detection of Plasmodium falciparum to examine the performance of this novel mobile application in detecting Plasmodium falciparum. 50 blood samples from suspected malaria patients were collected in EDTA tube from different hospitals in Lahore. The collected samples were screened for detection of Plasmodium falciparum by using Malaria screener and polymerase chain reaction (PCR). PCR method was considered as the reference method for the diagnosis of malaria. Out of 50 samples, 30 (60%) were identified as positive (P. vivax) by the analysis of blood smears by malaria screener and only 18 (36%) were identified positive for Plasmodium vivax and none was found positive for Plasmodium falciparum. The sensitivity and specificity of malaria screener were calculated and found to be 50% and 77.77%, respectively. Cohen's kappa coefficient was used to compare agreement between malaria screener and PCR, considered as reference standard. When comparing the malaria screener and the PCR, this study discovered that there was moderate agreement (k=0.49).

Veterinary Sciences and One Health Journal, 2023

Brucellosis is one of the most important zoonotic diseases worldwide. Brucellosis is not only haz... more Brucellosis is one of the most important zoonotic diseases worldwide. Brucellosis is not only hazardous to human but also causes a huge economic loss due to abortions in cattle, buffaloes, sheep and goats. Three main species are involved in brucellosis, i.e. Brucella melitensis in goats and sheep, Brucella suis in pigs, and in cattle Brucella abortus. The present study was designed to serologically diagnose and to find the associated risk factors of brucellosis in Punjab, Pakistan. In a total of buffaloes 100 and human 20 serum samples were randomly collected from different dairy farms, and were screened for brucellosis by Rose-Bengal Plate Test (RBPT). The positive RBPT samples were examined by indirect ELISA. Data of all animals was recorded on predesigned data captured form, which included age, sex, breed, housing system, feeding system, management, BCS, vaccination etc. Overall occurrence of Brucella abortus was recorded as 15 %(15/100) by RBPT and 5% by indirect ELISA in buffaloes. Moreover, in humans it was 5% (1/20) by RBPT and 0% by indirect ELISA. The higher occurrence was recorded in more than 5 years old age buffaloes, with larger herd sizes having an abortion history and retained placenta. This is the first study exploring buffaloes and human brucellosis in Punjab. The outcomes of this study will be helpful for farmers and veterinarians in evolving effective control plans.

Veterinary Sciences and One Health Journal, 2023

The study was conducted to evaluate efficacy of controlled internal drug releasing device (CIDR) ... more The study was conducted to evaluate efficacy of controlled internal drug releasing device (CIDR) and to assess levels of hormone i.e. Progesterone and minerals i.e. Calcium, Phosphorus during synchronization in postpartum anestrous Nili-Ravi buffaloes during low breeding season. For this purpose, a total of twenty buffaloes were divided into two groups each with 10 animal in each group. For the treatment group, CIDR was inserted intra-vaginal for 14 days and an injection of 2ml Dalmazine (150 µg d-cloprostenol) per animal was administered by intramuscularly on day 13. After CIDR removal, animals were observed for estrus and subsequently inseminated 12 and 24 hours after standing heat. Pregnancy was assessed at day 60 post insemination. In both groups, blood sampling was done at day 0, 7, 9, 11, 14, and 21 Post Artificial Insemination (AI) for analysis of Progesterone, calcium and phosphorus and difference was calculated between treatment and control group. Further, their difference was evaluated between pregnant and non-pregnant buffaloes. The progesterone concentration was measured through ELISA while levels of calcium and phosphorus were measured through spectrophotometry. The estrus intensity, ovulation rate and pregnancy rate were significant (P<0.05) higher in group A buffaloes. It was concluded that CIDR can be used efficiently for induction of estrus during low breeding season in buffaloes.

Veterinary Sciences and One Health Journal, 2023

In livestock, chronic diarrhoea is one of the clinical disorders associated with copper deficienc... more In livestock, chronic diarrhoea is one of the clinical disorders associated with copper deficiency. The purpose of this study will be to survey copper deficiency related chronic diarrhea in cattle and buffaloes in urban and peri-urban areas of Faisalabad. Study animals (n = 384) will be selected by unknown status of prevalence, the sample size was calculated by keeping the 50% expected prevalence with 95% confidence limit. A detailed proforma was designed and used to investigate the type of fodders and mineral supplements given to the animals in the study area. Blood samples were collected from animals suffering from chronic diarrhea in the study area. Sera samples were used to assess copper levels by atomic absorption spectrophotometry at Hi Tech Laboratory, University of Agriculture Faisalabad. Prevalence of chronic diarrhoea in cattle and buffaloes in the study area was calculated by simple binomial method and Chi square test were used to determine copper levels between diarrhoeic and non-diarrhoeic animals.

Veterinary Sciences and One Health Journal, 2023

N. caninum is an animal parasitic protozoan. Neospora can persist in the majority of dairy cows i... more N. caninum is an animal parasitic protozoan. Neospora can persist in the majority of dairy cows is facilitated by its vertical transmission. N. caninuminduced abortion in bovines is a significant contributor to economic losses in Pakistan's burgeoning dairy sector. Research comprises four brain samples of aborted bovine fetuses from several regions in the province of Punjab. The objective of this study was the isolation and propagation of the parasite, which is the first step toward the production of a vaccine against this parasite. Microscopic and molecular identification of N. caninum was performed through PCR. N. caninum was isolated from the brains of aborted fetal samples, propagated in the mice and then mice were killed to take brains, Neospora parasite was confirmed using PCR. Continuation of the experimentation meant for isolation and propagation by exploiting dams to be found seropositive in the planned studies, will be an important perspective of the study.

Veterinary Sciences and One Health Journal, 2023

Haemonchosis is responsible for a wide range of health problems, economic losses in small ruminan... more Haemonchosis is responsible for a wide range of health problems, economic losses in small ruminants and is characterized by anemia, dehydration, loss of appetite and body condition, retarded growth, and reduced production performance. Haemonchus Contortus has develops benzimidazole resistance. Total of 100 samples (n = 50 sheep, n = 50 goats) was collected directly from animals' abomasum. Collection of samples was done from the local abattoir of Shahpur Kanjra Lahore (Pakistan). The genomic DNA of adult male H. contortus was extracted by using DNA extraction kit and phenol chloroform method. Pair of primers was developed for H. contortus. After that, the PCR product was run on 1.2% agarose gel. The product was compared with known marker of DNA ladder to have correct amplified band. The PCR amplified band was purify by using gel extraction kit and sent with primers for sequencing. The sequences were aligned and analyzed by BIOEDIT software. By this alignment tool, mutations in different codon were observed and compared with β tubulin gene sequence from gene bank of NCBI. Significance of mutations at different locations was evaluated by using Chi Square Test. Data on mutation in H. contortus was analyzed by Pearson's chisquare test by using SPSS version 17. The results obtained showing 0.004% significance in sheep. While no mutation was observed in goats. That is no significant as compared to sheep samples. Percentages of negative and positive samples were calculated. Mutations in different codon in β tubulin gene were observed and their percentages were compared. By finding the mutations, this study helped to trace out the BZ's resistance in H. contortus in local sheep and goats and also determine the changes at codon 200 of sheep.

Veterinary Sciences and One Health Journal, 2023

Wild and domestic bovids are susceptible for obligate blood parasites consisting of Theileria, Ba... more Wild and domestic bovids are susceptible for obligate blood parasites consisting of Theileria, Babesia and Anaplasma. These are the most economically important and lethal parasites in dairy industry. The haemoparasites prevalence depends on managemental factors such as locality, herd size, tick control strategies, tick infestation rate and housing system which have been neglected. Therefore, the current study was designed to investigate the correlation of managemental practices with prevalence of haemoparasites. For this purpose, a total of 50 (30 before and 20 after medication) blood samples were collected from cattle of physically and clinically showing the symptoms of haemoparasites infection. Upon microscopic examination, the prevalence of theileriosis, babesiosis, anaplasmosis and mix infection was 40% (9/30), 30% (7/30) and 23.33% (5/30) and 20% (6/30) respectively before medication. After medication with buparvaquone and imidocarb dipropionate combined with oxytetracycline, the blood samples of those animals were again processed for microscopic examination. The prevalence of theileriosis, babesiosis and anaplasmosis was 30% (6/20), 20% (4/20) and 10% (2/20), respectively after 24-48 hour of treatment. The medication was effective to treat the infected animals but lacks managemental practices were the source of re-infection. It is concluded that only treatment of haemoparasites is not the way to control disease but necessary to improve the management eradicate the habitat of ticks.