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Tropical theileriosis is a lymphoproliferative disease caused by Theileria annulata and is transmitted by Ixodid ticks of the genus Hyalomma. It causes significant losses in livestock, especially in exotic cattle. The existing methods for controlling it, chemotherapeutic agents and a vaccine based on an attenuated schizont stage parasite, have several limitations. A promising solution to control this disease is the use of molecular vaccines based on potential immunogenic proteins of T. annulata. For this purpose, we selected five antigenic sequences of T. annulata, i.e. SPAG-1, Tams, TaSP, spm2, and Ta9. These were subjected to epitope prediction for cytotoxic T lymphocytes, B-cells, and helper T lymphocytes. CTL and B-cell epitopes with a higher score whereas those of HTL with a lower score, were selected for the construct. A single protein was constructed using specific linkers and evaluated for high antigenicity and low allergenicity. The construct was acidic, hydrophobic, and thermostable in nature. Secondary and tertiary structures of this construct were drawn using the PSIPRED and RaptorX servers, respectively. A Ramachandran plot showed a high percentage of residues in this construct in favorable, allowed, and general regions. Molecular docking studies suggested that the complex was stable and our construct could potentially be a good candidate for immunization trials. Furthermore, we successfully cloned it into the pET-28a plasmid and transformed it into the BL21 strain. A restriction analysis was performed to confirm the transformation of our plasmid. After expression and purification, recombinant protein of 49 kDa was confirmed by western blotting. An ELISA detected increased specific antibody levels in the sera of the immunized animals compared with the control group, and flow cytometric analysis showed a stronger cell-mediated immune response. We believe our multi-epitope recombinant protein has the potential for the large-scale application for disease prevention globally in the bovine population. This study will act as a model for similar parasitic challenges. [ABSTRACT FROM AUTHOR]

Bovine coronavirus (BCoV) is a major viral pathogen linked to respiratory and enteric problems in newborn calves. The goal of this study is to look into the molecular occurrence, haemato-biochemical changes, and risk factors associated with the occurrence of BCoV infection in cattle calves at different dairy farms and small households in the district of Jhelum, Pakistan. From July 2020 to June 2021, 200 faecal samples were collected from newborn cattle calves exhibiting symptoms of diarrhoea and dysentery. S&C Biotech Bovine Coronavirus Antigen Rapid Test Kits were used to screen samples, which were then subjected to RT-PCR for molecular characterization. For the haemato-biochemical analysis, blood was taken from calves infected with BCoV. A questionnaire was used to collect information about the risk factors associated with the occurrence of BCoV infection. BCoV infection was found in 3.5% (7/200) of calves using diagnostic screening kits and 3.0% (6/200) of calves using RT-PCR. Breed, age, sex, vomiting, previous history of diarrhoea, body conditions, food type, colostrum feeding, deworming history, living environment, seasonal variation, and cohabitation with other animals were all assumed risk factors for BCoV infection (p<0.05). There was a significant (p<0.001) decrease in MCV (fl) and a significant (p<0.006) increase in TLC (1x103/cm3) on haematological analysis in BCoV-infected calves. TEC (1x106/cm3), WBC (1x106/cm3), RBC (1x106/cm3), and Hct% were significantly (p<0.05) higher. Similarly, infected calves had a non-significant (p<0.090) increase in K (mEq/L) and a significant (p<0.000) decrease in Na (mEq/L), Ca (mmol/L), Cu (mol/L), and Fe (mol/L) on biochemical analysis. It was concluded that the occurrence of BCoV infection was predisposed by assumed risk factors, and haemato-biochemical alterations were observed in BCoV infected calves. [ABSTRACT FROM AUTHOR]

Mastitis is the most costly disease of dairy cattle and poses colossal economic losses to the farming community worldwide. The role of vaccination in controlling mastitis is of paramount importance and significantly reduces the use of antimicrobial agents. This study aimed at preparing and evaluating inactivated polyvalent mastitis vaccines containing bovine origin Staphylococcus aureus (tst), Streptococcus uberis (cpn-60 targeted STUB), and Escherichia coli (aggR) in a rabbit model. Four types of mastitis vaccines viz. formalin-inactivated plain polyvalent mastitis vaccine (FPPV), formalin-inactivated montanide adjuvanted polyvalent mastitis vaccine (FMPV), formalin-inactivated aluminum hydroxide adjuvanted polyvalent mastitis vaccine (FAPV), and formalin-inactivated montanide plus aluminum hydroxide adjuvanted mastitis vaccine (FMAPV) were prepared after confirmation for antigenicity and immunogenicity and evaluated for safety, side effects, challenge protection assay, sterility and humoral response in laboratory animals. Safety test showed no general adverse reactions to the vaccines when injected either into mice or rabbits or cow calves. Challenge protection studies revealed a significantly higher survival rate in vaccinated mice and rabbits compared to placebo groups. None of the vaccines when streaked onto culture media showed any growth indicating sterility of the vaccines. Repeated measures ANOVA was applied to compare mean O.D values of treatment groups for evaluation of humoral immunity. Serum ELISA O.D values against S. aureus (tst), Str. uberis (cpn-60 STUB) and E. coli (aggR) in vaccinated groups randomly inoculated i/m at thigh region @ 0.2 ml per rabbit were significantly higher (p<0.05) compared to control. Likewise, the humoral immune response of FAPV was highest followed in order by FMPV, FMAPV, and FPPV. It was concluded that newly-prepared polyvalent mastitis vaccines were safe to use, protective, and elicited a significant humoral immune response in rabbits. [ABSTRACT FROM AUTHOR]

Coccidiosis is a preeminent threat to productivity in broiler industry. This prospective study was undertaken to evaluate the anti-coccidial efficacy of guar (Cyamopsis tetragonoloba) and salinomycin sodium in broiler Chicken. One hundred and twenty broiler chicks were divided into four groups with each having 30 birds. Group A had salinomycin sodium supplementation, group B received Guar (Cyamopsis tetragonoloba) whereas groups C and D were positive and negative controls respectively. Salinomycin and guar (Cyamopsis tetragonoloba) were administered following oral inoculation of 50.000 oocysts of Eimeria specie in all birds except the ones inducted into negative control group. Weight gain, feed conversion ratio, Fecal oocysts and blood profile of these birds were determined weekly. When histopathology was performed only a couple of coccidian parasites were identified in the intestinal caeca of birds treated with guar, whereas none were seen in birds treated with salinomycin sodium. It was concluded that results in terms of weight gain and feed conversion ratio were more favorable in case of salinomycin supplementation. However, guar exhibited greater efficacy as an antiprotozoal agent with significant reduction in fecal shedding of oocysts. In contrast to groups receiving salinomycin, severity of bloody diarrhea and mortality rates amongst infected birds subjected to guar were appreciably lower as well. [ABSTRACT FROM AUTHOR]

Aflatoxins are the most deleterious fungal metabolites in humans, animals and plants. Recently, more attention has been paid to the occurrence and growth of Aspergillus spp. In this study, the effects of pH, light, relative humidity (RH) and temperature on the growth of Aspergillus spp. were investigated using broth medium. Results revealed highest mean of dried mycelial weight (365.67mg) at pH 4.0. The highest spores per ml (8.243 × 107) at pH 5.0. Similarly, growth during darkness was higher on first day of incubation in A. candidus and A. niger, while A. flavus and A. fumigatus showed the highest growth under continuous light. Lower RH% (32.5%) favored only the growth of A. niger. However, the growth in other Aspergillus species was significantly increased by RH% (50.5%, 85.0% and 100%). At 40C, only A. fumigatus and A. flavus showed significant (P<0.05) growth. There were significant differences in all the treatments (P<0.05). This proposes that these interacting environmental conditions impact significant effect on Aspergillus growth. [ABSTRACT FROM AUTHOR]

Introduction: Mycobacterium tuberculosis is the main cause of tuberculosis in humans, accounting for numerous illnesses and thousands of fatalities globally. Data regarding the association of various risk factors and TB in livestock farmers in Pakistan is scarce. Methodology: A retrospective matched case-control study of TB cases was performed in Lahore, Pakistan to investigate the potential risk factors that lead to the development of TB in Pakistani livestock farmers. A total of 170 participants were included in the study. The case was matched with control based on neighborhood and the case-control ratio was kept 1:1. Data were statistically analyzed using R version 4.2.1. Conditional logistic regression was conducted to identify biologically and statistically plausible risk factors associated with the TB outcome among livestock farmers. Results: In univariable analysis, 10 risk factors were identified (p < 0.05). Gender, age, being married, family type, living in a big family, BCG vaccination status, history of smoking, working at a cattle farm, co-housing with cattle at night,, consumption of raw milk. The multivariable model identified four risk factors i.e., consumption of raw milk (Odds Ratio [OR]: 7.7; 95% Confidence Interval [CI]: 1.95-30.68), living in big family (OR: 6.2; 95% CI: 1.25-30.82) and working at cattle farm (OR: 4.2; 95% CI: 1.08-16.56), while gender was found to be a protective factor with OR < 1 (OR: 0.06; 95% CI: 0.01-0.26). Conclusions: This study demonstrated that sociodemographic risk factors and exposure to infected cattle can influence the development of TB in farmers. [ABSTRACT FROM AUTHOR]

Infectious bovine keratoconjunctivitis (IBK) commonly known as pinkeye is a contagious non-fatal bacterial infection of the eye that has tremendous economic impact on the dairy industry. Nevertheless, it is a much-neglected dairy issue in Pakistan. The current study was aimed to find out the prevalence and risk factors associated with pinkeye infection in commercial dairy herds in Pakistan. This study recruited 10,573 cows in 20 commercial dairy herds holding ≥25 cows per herd. The cows were assessed based on eye lesions using a scale from 1 to 4, with 4 being the most severe of all. Culture of subconjunctival swab samples was also undertaken. The overall prevalence of pinkeye was 6.86% with Holstein-Friesian breed being the most affected (7.25%) followed by crossbred (6.89%) cattle. Cow hygiene and farm hygiene scores were indirectly related to the prevalence of pinkeye. Prevalence was also significantly (P<0.05) associated with seasons of the year, being highest in humid summer months while lowest in winter. The prevalence of pinkeye was significantly higher (<0.0001) in cows exposed to sunlight than unexposed ones. Cows on farms performing farm disinfection occasionally had 1.52 times higher odds of being diseased than cows on farms with once-a-week disinfection. Likewise, cows on farms using no fly control program had 2.8 times higher likelihood of pinkeye than those on farms having fly control program. In addition, prevalence of pinkeye was significantly higher (P<0.05) in cows with low body condition scores. It was concluded that Pinkeye is a significant health concern in commercial dairy herds in Pakistan and the risk factors influencing the prevalence of this disease may include cow hygiene, farm management practices, and the environment. [ABSTRACT FROM AUTHOR]

Introduction: The main objective of the study was to estimate the burden of occupational tuberculosis infection in high-risk occupational workers and to identify risk factors associated with the prevalence of Mycobacterium tuberculosis complex (MTBC). Methodology: An analytical cross-sectional study was conducted among high-risk occupational workers including veterinarians, abattoir workers, animal handlers, livestock farmers, and microbiology laboratory workers. Sputum samples were collected from 100 participants and polymerase chain reaction (PCR) tests were done to diagnose tuberculosis (TB) infection. Data on potential risk factors was collected in a pre- designed questionnaire. The MTBC prevalence ratio was estimated. Logistic regression analysis was conducted to identify risk factors and the crude odds ratio (OR) was calculated. Results: Among the 100 enrolled high risk occupational workers, the prevalence of MTBC was 46% (95% CI: 35.98--56.25). Living in a joint family (OR 3.85, 95% CI: 1.58--9.37), and use of unpasteurized milk (OR 3.42, 95% CI: 1.4--8.39), were significantly associated with MTBC infection. Conclusions: Tuberculosis is a significant health burden in high-risk occupational groups, especially animal handlers and laboratory workers, in Lahore, Pakistan. The study also emphasized the need for formal work-related training, and enhanced zoonotic TB awareness among occupational workers. [ABSTRACT FROM AUTHOR]

Mastitis is a complex and multifarious disease that adversely affects the physico-chemical properties, composition and yield of milk and risks of premature culling of affected animals. A total of 320 Cholistani cattle were screened for subclinical mastitis using California mastitis test in and around district Bahawalpur. The blood and milk samples were only collected from subclinical infected cattle. Milk samples obtained from cattle with subclinical infection revealed significantly (P<0.05) negative correlation of Na with K, Ca, Mg, P, Fe and Zn while significant (P<0.05) positive correlation with aspartate aminotransferase, lactate dehydrogenase and alkaline phosphatase. Serum samples from infected animals revealed significant (P<0.05) positive correlation of Ca with P and Zn while significant negative correlation of Zn and P with enzymes. Milk samples from non-infected cows showed a significant (P<0.05) negative correlation between Ca and K while positive correlations between Ca and P. Blood samples from non-infected animals revealed significant positive correlation between zinc, AST and ALP while significant (P<0.05) positive correlation between LDH, ALP and AST. From the present study, it was concluded that correlation of milk and blood electrolytes, trace minerals and enzymes are suitable diagnostic technique to detect sub-clinical mastitis. [ABSTRACT FROM AUTHOR]

The aim of the present study was to determine the deleterious effects of prolonged oral cyanide insult on various organs and tissues in rabbits. For this purpose, 12 locally bred adult male rabbits were allocated into two groups of 6 viz. control and experimental. Rabbits in control group were offered feed only while the rabbits in experimental group received feed plus potassium cyanide (KCN) at 3 mg/kg body weight orally for a period of 40 days. None of the rabbit in both the groups demonstrated any of the gross changes in any organ on postmortem examination. Liver was normal in size, shape, texture and color. Kidneys were also normal in size and color. Histopathological examination revealed severe hepatocyte vacuolation and degeneration in liver of rabbits in experimental group. There was also excessive congestion in liver and bile duct of rabbits in experimental group. Kidneys of rabbits in experimental group demonstrated severe glomerular and tubular necrosis and congestion. In the tubular epithelial cells, pyknotic nuclei were also present. On the other hand, heart and pancreas of rabbits in both control and experimental group did not show any histopathological change in microscopic structures. In conclusion, prolonged oral cyanide administration could have harmful effects on liver and kidney functions. [ABSTRACT FROM AUTHOR]

The objective of the study was to determine the effect of dietary aflatoxin B1 (AFB1) in growing male buffaloes in terms of feed intake, hematological and serum biochemical parameters. Thirty six bovine growing male buffaloes between 6 months to 12 months of age were divided into four groups to receive diet having no AFB1, 0.6mg/kg, 0.8mg/kg and 1.0mg/kg AFB1 of feed per animal per day for 28 days. The results indicate that the average daily feed intake of AFB1-treated growing male buffaloes significantly declined (P < 0.05) compared with the control. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and creatinine were significantly increased (P < 0.05) in response to AFB1, However change in blood urea nitrogen (BUN) levels was non-significant values of all hematological parameters decreased significantly (P < 0.05) in1.0mg/kg AFB1 group as compared to other groups after day 14 onward. [ABSTRACT FROM AUTHOR]

This study was aimed to compare the efficacy of aqueous garlic extract, sodium nitrite (SNT), sodium thiosulfate (STS) and hydroxocobalamin against oral cyanide exposure in rabbits. For this purpose, forty two adult male rabbits were divided randomly into 7 groups of 6 animals (A-G) each. Rabbits in group A were offered feed only and served as negative control, while the rabbits in group B received feed plus potassium cyanide (KCN) at 3mg/kg orally and were kept as positive control. Animals in group C received feed, KCN and intraperitoneal injection (IP) of aqueous garlic extract at 500mg/kg. Rabbits in group D were given feed, KCN and IP injection of STS at 600mg/kg. Members in group E received feed, KCN and IP injection of both aqueous garlic extract at 500mg/kg and SNT at 20mg/kg. Animals in group F were given feed, KCN and IP injection of both STS at 600mg/kg and SNT at 20mg/kg, while the rabbits in group G received feed, KCN and IP injection of hydroxocobalamin at 300mg/kg. The treatments were given to respective groups for 40 days. The efficacy of the antidotes was measured on the basis of changes in biochemical profile of rabbits in each group. In this study, hydroxocobalamin was found to be significantly more effective cyanide (CNI) antidote than garlic, STS, SNT plus garlic extract, or SNT and STS, either alone or in combination. A combination of SNT and garlic extract was the second most effective CNI antidote. The efficacy of garlic alone was significantly higher than STS alone or in combination with SNT. The efficacy of combined SNT and STS was superior to STS alone in treating rabbits with CNI toxicity. In conclusion, aqueous garlic extract alone or in combination with STS can effectively be used against cyanide toxicity. [ABSTRACT FROM AUTHOR]

Bovine mastitis causes severe economic losses in terms of poor milk production and culling of chronically infected animals. In present study milk samples were collected from Cholistani cattle and examined using California mastitis test (CMT). Different milk and blood serum biochemical parameters were investigated. The results indicated that milk pH, milk electrical conductivity, malondialdehyde concentration and total dissolved solids were significantly (P<0.0001) increased with increase in CMT score. The values of milk fat, protein, lactose and solids not fat were significantly reduced in milk samples of sub-clinically infected cattle. The results showed that the values of total milk leukocyte and neutrophil counts were significantly higher in infected cattle, while the percentages of monocytes, macrophages and lymphocytes were lower. The levels of potassium, calcium, magnesium, phosphorous, iron and zinc were significantly lower in milk and blood serum of infected than normal cattle. The enzymes such as aspartate aminotransferase, lactate dehydrogenase and alkaline phosphatase were higher in milk and blood serum of infected animals. Similarly, malondialdehyde and nitric oxide were significantly higher in infected animals. The results showed that mastitis poses deleterious effects on milk and blood of infected animals. [ABSTRACT FROM AUTHOR]

The current study was planned to investigate the potential risk factors and their association with mastitis in Cholistani cattle. For this purpose milk samples were collected and examined for mastitis using California Mastitis Test. The results showed an overall 21.96% prevalence of subclinical mastitis. The analysis of variance did not show significant association of tail size, live body weight, mid and base teat diameter with mastitis. The analysis of variance showed significantly higher involvement of right rear quarters. Results showed that teat length, teat end to floor distance and milk yield (P<0.001) were significantly lower in mastitic animals while teat apex diameter, parity and udder depth was significantly higher (P<0.001). The frequency analysis revealed significant association of mastitis between different groups such as age, udder depth, lactation stage, parity, teat shape, udder shape, teat lesions and live body weight (P<0.001). Bivariate logistic regression analysis indicated significant association of age, parity, lactation stage, teat shape, udder depth, teat and udder lesions, teat end to floor distance, milking technique, milk leakage and type of floor with mastitis. [ABSTRACT FROM AUTHOR]

Twelve adult rabbits bred locally were divided into two equal groups of 6; experimental and control groups. Rabbits in the experimental group were orally dosed with KCN at 3mg/kg body weight for 40 consecutive days. Members in control group were given placebo (distilled water) for the same period. Animals in both groups were offered feed at 90gm/kg/day while ample drinking water was available ad lib. Feed consumption and body weight of rabbits in both the groups were recorded. Blood samples were also drawn to determine various hematological parameters. Statistical analysis revealed a non-significant difference of total and daily feed intakes in rabbits of experimental and control groups. Whereas the feed efficiency of rabbits in the experimental group were significantly reduced (P<0.05) compared to controls. Likewise a significant decrease in body weight gain of rabbits in experimental group (P<0.05) was observed. A non-significant difference (P>0.05) was observed in leukocyte count, differential leukocyte count and platelets of rabbits in both the groups. Erythrocyte count, hemoglobin concentration, packed cell volume and mean corpuscular hemoglobin were significantly decreased in treated rabbits. It was concluded that chronic cyanide intake had a deleterious effect on feed efficiency, growth rate and blood components of rabbits. [ABSTRACT FROM AUTHOR]

The article discusses a study which determined the infection rate of theileriosis in Lohi sheep and investigated an economical and effective remedy for this protozoal problem in Okara District, Pakistan. The study also examined the effect of this disease on various blood parameters. The study found that 16.5% sheep were positive of which 65.1% were clinically symptomatic and showed schizonts of thielerial parasite in their lymphocytes on the basis of Giemsa stained blood smears. Buparvaquone, however, was found to be better than oxytetracycline in treating the disease.

Coccidiosis is a protozoal and occasionally fatal diarrheic disease of goats imposing heavy economic losses tofarming community. This study aimed to evaluate the efficacies of Furazolidone, Sulfadimidine and Amprolium against coccidiosis in Beetal goats. Twenty-four (24) Beetal goats naturally infected with coccidiosis were randomly divided intofour groups of 6 (A-D). Goats in groups A, B and C were treated orally with Furazolidone (10mg/Kg), Sulfadimidine (100mg/Kg) and Amprolium (55mg/Kg), respectively for 7 days. Goats in-group D served as positive control. Oocysts per gram (OPG) of feces counts of individual goats in each group were performed on Days; 0 (pre-treatment) 7, 14 and 21 (post-treatment). OPG counts amongst goats in all groups at day 0 were not significant (P>0.05). On days 7, 14 and 21, OPG values decreased significantly (P<0.05) in groups A, B and C compared to group D. The efficacy of Furazolidone, Sulfadimidine and Amprolium was 98.6, 98.0 and 99.6 percent, respectively on Day 21 (end of trial). Statistically, the efficacies of three drugs were not significantly different (P>0.05). In conclusion, Furazolidone, Sulfadimidine and Amprolium are well-tolerated and any one of these may be recommended to effectively treat coccidiosis in Beetal goats. [ABSTRACT FROM AUTHOR]

The current study aimed to investigate the prevalence and molecular characterization of biofilm-positive S. aureus isolates from bovine subclinical mastitis. The study also highlights the role of commonly used NSAIDs and ivermectin to modulate the S. aureus-associated antibiotic resistance. The results found a 41.41% S. aureus prevalence, out of which 25.79% isolates were biofilmpositive based on Congo red agar, microtitre plate test, and presence of icaA gene. Phylogenetic analysis of study isolates showed a high similarity with Egyptian and Indian icaA-positive S. aureus isolates. The comparative antibiotic resistance profiling showed a significantly (p<0.05) higher resistance to gentamicin, oxytetracycline, and cotrimoxazole by biofilm-positive isolates compared to nonbiofilm forming isolates. The prevalence of methicillin and vancomycin resistant S. aureus was 62.5 and 20.83%, respectively. Antimicrobial effects of nonantibiotics against study isolates accessed through well diffusion method showed higher zones of inhibition for meloxicam followed by flunixin, ketoprofen, and ivermectin. The combinations of resistant antibiotics with non-antibiotics were investigated using well diffusion method and checkerboard assay. The combinations of amoxicillin/meloxicam, cotrimoxazole/flunixin, cotrimoxazole/ketoprofen, and gentamicin/flunixin on well diffusion method and cotrimoxazole/flunixin, amoxicillin/ketoprofen and gentamicin/flunixin on checkerboard assay revealed synergistic interactions. The study concluded that biofilm positive S. aureus is an emerging and prevailing cause of bovine mastitis in dairy farms of Pakistan. The increasing antibiotic resistance in S. aureus can be modulated by combining the resistant antibiotics with NSAIDs, especially flunixin and ketoprofen. [ABSTRACT FROM AUTHOR]

Indiscriminate use of antibiotics to treat Staphylococcus aureus bovine mastitis may impair mammary glands immune function and develop bacterial resistance. Consequently, developing alternative remedies to treat mastitis is imperative. Current study evaluated plain multidrug resistant S. aureus vaccine (PMRSAV), Montanide oil adjuvanted multidrug resistant S. aureus vaccine (MMRSAV), and Aluminum hydroxide adjuvanted multidrug resistant S. aureus vaccine (AMRSAV) in lactating dairy cattle. Serum samples through IHA revealed that MMRSAV provoked maximum antibody titer at day 90 (274.4±0.41) whereas AMRSAV (147±0.46) and PMRSAV (78.8±0.44) produced highest antibody response at day 60. The cumulative mean serum IHA antibody titer was recorded highest for MMRSAV (148.4) than AMRSAV (78.4) and PMSAV (34.1). The peak milk whey antibody titer was seen at day 60 for PMRSAV (9±0.23) and AMRSAV (14.7±0.24) whereas it was observed highest at day 90 in MMRSAV (26.2±0.25). Somatic cell count (SSC: × 105 mL-1) observed decline trend in all the vaccinated cattle until day 90 for PMRSAV (1.449±0.219), day 120 for MMRSAV (1.201±0.097) and AMRSAV (1.327±0.104). During study period, the highest quarter-based incidence was observed in control group (25%) followed by PMRSAV (17.5%), AMRSAV (10%) and MMRSAV (5%) resulting incidence reduction (80%) for MMRSAV followed by AMRSAV (60%) and PMRSAV (30%). Post challenge lowest overall mean somatic cell count was recorded in group vaccinated with MMRSAV (2.323±0.46) followed by AMRSAV (3.006±0.43), PMRSAV (3.759±0.82) and unvaccinated control group D (7.798±1.11). The study concluded that MMRSAV is the most effective in preventing multidrug resistant S. aureus mastitis in dairy cattle. [ABSTRACT FROM AUTHOR]

Background and Aim: Anaplasma marginale is an obligate intraerythrocytic rickettsial parasite that infects cattle in tropical and subtropical regions. There is no evidence that A. marginale inoculation can be used to culture Rhipicephalus annulatus in rabbits. This study aimed to determine the molting of R. annulatus larvae, nymphs, and adults on rabbits as well as nymphs and adults of R. annulatus on calves with or without A. marginale. Transstadial, horizontal, and transovarial transmissions of A. marginale in R. annulatus reared on rabbits and calves were evaluated. Materials and Methods: Engorged female ticks were collected from field samples of A. marginale-infected and noninfected cattle. We divided the eight rabbits into two groups: A and B. Group A rabbits were infected with A. marginale through parenteral inoculation, whereas Group B rabbits were kept as a control. The "clean rabbits" in Group B were observed for tick rearing without A. marginale. Polymerase chain reaction was used to screen A. marginale in rabbits and stages of tick. The complete life cycle of R. annulatus with or without A. marginale was observed on rabbits. Results: A 6.5-day longer life cycle was observed in ticks harboring A. marginale than in ticks without A. marginale. To observe transstadial transmission, transstadial, horizontal, and transovarial transmissions of A. marginale in R. annulatus ticks were experimentally observed in one clean calf fed separately with infected nymphs and female adult ticks. Conclusion: We experimentally observed transovarian, transstadial, and transovarial transmission of A. marginale in R. annulatus ticks as a biological vector reared on calves and rabbits. We used rabbits as a model animal for rearing R. annulatus ticks and culture of A. marginale. [ABSTRACT FROM AUTHOR]

The emergence and inter-species dissemination of new tick-borne pathogens have become a serious health hazard in domestic animals. Ehrlichiosis caused by Ehrlichia (E.) ruminantium in small ruminants is becoming an emerging and prevailing issue adversely affecting the health and production of sheep and goats. The current study was designed to investigate the Ehrlichia infection load in the sheep and goat population of South Punjab, Pakistan. The phylogenetic analysis of local isolates along with risk factor analysis and comparative hematological analysis was also conducted. A total of 192 blood samples consisting of sheep (n=96) and goat (n=96) were subjected to microscopy and PCR. The study results revealed an overall prevalence of 04.69 and 7.81% based on microscopy and PCR respectively. A higher molecular prevalence of Ehrlichia was observed in goats (9.38%) as compared to sheep (6.25%). The phylogenetic tree constructed by the maximum likelihood method exhibited the identity of study isolate with E. ruminantium isolated from various countries. The risk factor analysis showed a significant association of previous tick history, tick infestation, presence of shrubs, and tick control strategies adaptation with the disease occurrence. The hematological analysis observed a significant reduction in RBCs, hemoglobin, and hematocrit in infected animals compared to healthy animals. The study concludes that Ehrlichia infection is an emerging issue of small ruminants. The hematological findings can be helpful in early detection of disease in field conditions but molecular confirmation by PCR is necessary for accurate diagnosis of Ehrlichia infection while risk factor analysis will provide the relevant information to develop control measures for preventing the spread of disease. [ABSTRACT FROM AUTHOR]

Sub-clinical mastitis has remained the leading cause for decline in production of dairy animals as a silent epidemic all the times in Pakistan. A questionnaire-based cross-sectional study was conducted to assess the prevalence and potential risk factors for sub-clinical mastitis in the Pothohar region of Pakistan in 2018. Total of 104 cattle milk samples were collected from commercial and subsistent dairy farms. CMT positive samples were cultured and biochemical tests were conducted before confirmed on PCR for mecA gene Methicillin Resistant Staphylococcus aureus (MRSA) bacteria. In-vitro antibiotic susceptibility was also assessed. An overall prevalence of 71.1% was found; where cross-bred was found more susceptible (80.7%) as compared to other breeds. MecA gene-MRSA prevalence based on PCR was 54%. On regression analysis the potential risk factors identified here included; daily milk yield, parity, udder shape, teat morphology, shed type, quarantine of new animals and deworming of animals (OR>1; P-value<0.05). In MRSA confirmed isolates, Penicillin group was found highly resistant (92.5%) amongst all the groups. While amongst individual antibiotics Amoxicillin was found highly (100%) resistant followed by Cefixime and Spectinomycin. Whereas based on sensitivity Quinolone (Moxifloxacin 95%) group was the most sensitive (91.7%) followed by Sulphonamides (Sulphaphenazole 87.5%) and Amino-glycoside (Gentamycin 90%). An emerging pattern of mecA gene MRSA was recorded here with alarming sub-clinical mastitis prevalence in the study area. Immediate preventive measures need to be taken to address the problem. The findings of the current study will assess in control and prevention of subclinical mastitis in Pakistan. [ABSTRACT FROM AUTHOR]

Tropical theileriosis is a lymphoproliferative disease caused by Theileria annulata and is transmitted by Ixodid ticks of the genus Hyalomma . It causes significant losses in livestock, especially in exotic cattle. The existing methods for controlling it, chemotherapeutic agents and a vaccine based on an attenuated schizont stage parasite, have several limitations. A promising solution to control this disease is the use of molecular vaccines based on potential immunogenic proteins of T. annulata . For this purpose, we selected five antigenic sequences of T. annulata , i.e. SPAG-1, Tams, TaSP, spm2, and Ta9. These were subjected to epitope prediction for cytotoxic T lymphocytes, B-cells, and helper T lymphocytes. CTL and B-cell epitopes with a higher score whereas those of HTL with a lower score, were selected for the construct. A single protein was constructed using specific linkers and evaluated for high antigenicity and low allergenicity. The construct was acidic, hydrophobic, and thermostable in nature. Secondary and tertiary structures of this construct were drawn using the PSIPRED and RaptorX servers, respectively. A Ramachandran plot showed a high percentage of residues in this construct in favorable, allowed, and general regions. Molecular docking studies suggested that the complex was stable and our construct could potentially be a good candidate for immunization trials. Furthermore, we successfully cloned it into the pET-28a plasmid and transformed it into the BL21 strain. A restriction analysis was performed to confirm the transformation of our plasmid. After expression and purification, recombinant protein of 49 kDa was confirmed by western blotting. An ELISA detected increased specific antibody levels in the sera of the immunized animals compared with the control group, and flow cytometric analysis showed a stronger cell-mediated immune response. We believe our multi-epitope recombinant protein has the potential for the large-scale application for disease prevention globally in the bovine population. This study will act as a model for similar parasitic challenges., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Abid, Khalid, Suleman, Akbar, Hafeez, Khan and Rashid.)

Multi-drugs resistant (MDR) Staphylococcus aureus is creating challenges to cure cow mastitis, resulting in massive economic loss globally. It necessitates the adoption of prevention and control systems such as vaccination. Plain (PMRSAV), Montanide oil adjuvanted (MMRSAV) and Aluminum hydroxide adjuvanted (AMRSAV) vaccines were prepared using a molecularly characterized isolate of MDR S. aureus from bovine origin. Immunogenicity of the selected isolate was evaluated in five groups of rabbits (A-E) at different concentrations by measuring GMT via IHA from serum samples after booster shot. The group E provoked significantly higher (P<0.05) antibody titer with peak at day 28 (64±0.5) and cumulative mean antibody titer (CMT) of rabbits was highest (45.6) followed by groups C (35.9), D (32.7), B (30.3) and A (24.5). The concentration yielding maximum antibody titer was used for vaccines preparation. Vaccines were evaluated in different rabbits groups by inoculating PMRSAV, MMRSAV, AMRSAV and Placebo. Serum samples evaluated through IHA revealed that rabbits injected with MMRSAV produced highest antibody titer reaching its peak at day 45 (90.51±0.23) with a slight decrease until day 60 (80.63±0.17) followed by AMRSAV and PMRSAV. Challenge protection assay revealed the survival rates of rabbits in groups PMRSAV, MMRSAV, AMRSAV and Placebo as 83.3%, 100%, 100% and 16.7%, respectively. The study concluded that MMSAV and AMSAV were safe, efficacious and immunogenic in experimental rabbits.