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This study surveyed the existence of the Coxiella burnetii in ticks removed from camels in three different regions of the Punjab, Pakistan. A total of 325 ixodid ticks were analyzed for the occurrence of C. burnetii DNA by real-time polymerase chain reaction. Before analyses, all ticks were pooled into 30, 20 and 15 pools from Southern, Northern and Central Punjab, correspondingly. Each pool was comprised of five engorged or semi-engorged ticks according to species. Pooled based prevalence in Southern Punjab was eighteen from 30 pools (60%), leading to maximum-likelihood of true prevalence of 0.56% (95% CI 0.23-1.31), while pooled prevalence in Northern Punjab was eight from 20 pools (40%), leading to maximumlikelihood of the true prevalence of 0.44% (95% CI 0.31-0.89) and none of the pooled samples from Central Punjab were found positive for C. burnetii. During classification of tick pools based on species, infection was diagnosed in 75% (09/12) of H. dromedary, 42.8% (03/07) of H. anatolicum, 50% (01/02) of H. scupense, 60% (03/05) of R. microplus, and 50% (02/04) of R. annulatus in Southern Punjab. In comparison, infection was detected in 37.5% (03/08) of H. dromedary, 23% (01/04) of H. anatolicum, 0% of H. scupense, 66.7% (02/03) of R. microplus and 66.7% (02/03) of R. annulatus in Northern region. The detection of C. burnetii implies important role of ticks in the dissemination of this bacterium in Punjab, Pakistan and reveals that ticks have a major contribution to the epidemiology of coxiellosis in this ecological system. These judgments warrant further molecular investigations for better perception about C. burnetii epidemiology and its contributions to humans and animal disease in Pakistan. [ABSTRACT FROM AUTHOR]

Glanders is highly contagious and zoonotic bacterial disease of equines caused by Burkholderia mallei. Since many decades, glanders is endemic in eastern parts (Punjab) of Pakistan and has no information of disease in the north-western region (Khyber Pakhtunkhwa – KPK). Therefore, present study was aimed to determine the seroprevalence and associated risk factors in two draft equine populated districts (Charsada and Peshawar) of KPK. A total of 393 equine samples originating from different localities of study area were investigated for anti-B. mallei antibodies (glanders) using indirect enzyme linked immunosorbent assay (ID Vet, Grabels France). The overall seroprevalence was found to be 3.1% (95% CI: 1.6-5.3) which remained insignificantly (χ2 0.421, P=0.516) between the two districts. In Charsada, prevalence estimates varied significantly (χ2 11.986, P=0.017) among 5 localities and found insignificant (χ2 1.287, P=0.732) in different sampling areas of Peshawar district. Donkeys were more seropositive (4.1%; 4/195) than horses (2.1%; 8/194) which did not differ statistically (χ2 1.397, P=0.237) and all mules were tested negative. Prevalence slight increased with age of animals (>10 years) and with poor body conditions. Donkeys were more test positive (OR: 2.05, 95% CI: 0.61-6.94) than horses and similarly animals managed on communal feeding (OR: 1.84, 95% CI: 0.23-14.5) and water troughs (OR: 1.65, 95% CI: 0.21-13.04) had more chances of contracting glanders. Multivariable model indicated donkeys (OR: 4.58, 95% CI: 1.0, 20.9) and equines above 10 years of age (OR: 3.10, 95% CI: 0.7-13.3) were more likely to test positive for glanders. In conclusion, present study provides prevalence estimates of equine glanders in KPK regions, which appears to be an area of endemicity. Future large-scale investigations are seriously warranted in other equine populated districts before implementing an effective control plan for glanders. [ABSTRACT FROM AUTHOR]

Mastitis in goats, analogous to dairy cattle, is among the momentous diseases of economic value in all parts of the world. Local information of epidemiology of goat mastitis is scanty. In the present study, considering geographical distribution of Beetal goat, 172 flocks were randomly pulled out by “Survey tool box” to reduce biased sampling from perie-urban and urban areas of six districts of Punjab. By following the proportionate sampling strategy and assuming the highest population (70%) of Beetal all over the Punjab, Beetal strains were screened for the prevalence of mastitis from Faisalabad, Jhang, Chiniot, Bahawalpur, Muzzafargarh and Rajanpur districts. Screening of the flocks with surf field mastitis test indicated that overall prevalence of caprine mastitis in Beetal breed was 17.39% while in Beetal Faisalabadi strain 16.53% from 116 flocks, in Beetal Makhicheeni 18.06% from 32 flocks and in Beetal Nuqri 19.33% from 24 flocks. Microbiological examination of milk samples showed various types of mastitic microorganisms. The predominant isolates included Staphylococcus (S). hyicus (45%), S. xylosus (19%), S. simulans (16%), S. aureus (9%) and unidentifiable Staphyloccocal species (11%). Antibiotic sensitivity test was conducted on the most prevalent isolates. S. aureus and S. hyicus showed sensitivity to eight antibiotics: amoxicillin, ampicilin, lincomycin, sulfamethoxazole+trimethoprim, novobiocin, enrofloxacin, amoxicillin+ clavulanic acid and oxytetracycline while S. xylosus was sensitive to all antibiotics except novobiocin and S. simulans was sensitive to lincomycin, sulfamethoxazole+ trimethoprim, enrofloxacin and resistant to all other antibiotics. The study found over all 16.74% mastitis in different strains of Beetal. The most important parameters as; type of housing, farm hygiene, age, stage of lactation, parity, color, body condition score, teat ends, SFMT score and decreased milk yield were found pertinent risk factors so that prompt attention required for mastitis control in goats. [ABSTRACT FROM AUTHOR]

The present study was conducted to determine the sero-prevalence of Q fever and associated risk factors in institutional dairy farms in Punjab, Pakistan. A total 11 dairy farms were investigated from different areas of Punjab province. Out of these farms, a total of 827 animals (cows n=419 and buffaloes n=408) were investigated. Presence of anti-Coxiella burnetii antibodies was determined by indirect ELISA. The overall prevalence in dairy animals (cattle and buffaloes) was 6.1% (95% CI: 4.5-7.9). In cattle prevalence was higher (7.6%; 95% CI: 5.3-10.6) than in buffaloes (4.4%; 95% CI: 2.6-6.9). The probability of occurrence of disease in cattle was higher (7.63%; OR=1.9, 95% CI: 0.99-3.25) than in buffalo (1.00%; OR=1.00) The difference in sero-prevalence between the two-dairy species was significant (p=0.052, x² = 3.786). Stratification of Q fever as per the breed of cattle indicated that the sero-prevalence was the highest (13%) in Cholistani followed by Sahiwal (7.5%) and cross bred (2.6%). Animals age between 2.1 to 5 year (OR=2.56, 95% CI 1.05, 6.20), contact with small ruminants (OR=1.43, 95% CI 0.63, 3.25), tick infestations (OR=4.91, 95% CI 2.40, 10.00), dry-dusty environment (OR=1.55, 95% CI 0.54, 4.43), retained fetal membranes (OR=1.68, 95% CI 0.94, 5.73) and abortion history (OR=2.17, 95% CI 0.88, 5.35) were risk factors for the seropositivity of Q fever. The results of the study indicated that Q fever is prevalent in cattle and buffalo managed at institutional dairy farms in Punjab. [ABSTRACT FROM AUTHOR]

Echinococcus granulosus (G1/G3) and E. ortleppi (G5) are prevalent worldwide. Numerous studies have been conducted to characterize these parasites and utilized mainly cox1, nad1, and nad5 genes. However, there is a lack of comprehensive global data on genetic diversity in E. granulosus and E. ortleppi based on cox2 and nad6 genes. The current pilot research work was executed to look into the genetic variation of the E. granulosus (G1/G3) and E. ortleppi (G5) populations in Pakistan using cox2 and nad6 and infer phylogenetic relationships with other E. granulosus sensu lato populations. Followed by cyst collection from slaughtered cattle in Pakistan and DNA extraction, the full mitochondrial cytochrome oxidase 2 and NADH dehydrogenase subunit 6 were amplified. After confirmation of the identity of isolates through BLAST analysis, population indices pertaining to neutrality and diversity were calculated using the DNASp program. MJNs were constructed using PopArt for intraspecific data investigation. Nucleotide sequences (n=17) analyzed in this investigation were matched with GenBank reference sequences and Echinococcus genotypes G1 (n=5) and G3 (n=7) were discovered. Five sequences were identified as E. ortleppi (G5). For E. granulosus (G1/G3), a high haplotype diversity (Hd=1.00) was found for cox2 (582 bp), nad6 (456 bp), and concatenated cox2-nad6 (1038 bp) gene sequences. The overall value of Tajima's D for the entire population was negative and insignificant. Regarding E. ortleppi, cox2--nad6 π and Hd were 0.07290 and 1, respectively with insignificant Fu's Fs. A dataset of 40 cox2 and 35 nad6 sequences was utilized to form a medianjoining network to compare the geographical kinship between the Pakistani isolates and sequences reported elsewhere. The analysis of concatenated cox2-nad6 obtained in this study revealed a high haplotype diversity (1.00) while low nucleotide diversity (0.08207) was observed. Non-significant values of Tajima's D = -0.54361 and Fu's Fs = 1.303 were observed. High haplotype diversity in the current study shows a significant level of genetic divergence across haplotypes. The amalgamation of low π and high Hd indicates population expansion. We recommend amplifying a greater number of isolates in the future to ascertain the resolution power of cox2 and nad6 genes in the context of genotype differentiation. [ABSTRACT FROM AUTHOR]

Rodents are one of the most abundant and successful mammals on the planet. The rats harbor the metacestode stage of Hydatigera taeniaeformis serving as the intermediate hosts, whereas cats act as definitive hosts of the parasite. The incidence of the larval stage in rats has been documented in various regions of the world. To best of our knowledge, the mainstream studies on the genetic diversity of H. taeniaeformis are based on the cox1 gene which precludes an in-depth analysis of genetic variation in the parasite. To provide insights about nad1 gene-based genetic variation, in continuation of our previous work, we thereby in this study report H. taeniaeformis infection in the urban murine population from the Faisalabad district of Pakistan and presented genetic polymorphism using the nad1 gene. Out of 38 isolates investigated in the study, a total of 13 haplotypes were found with high haplotype diversity (Hd = 0.909), while the nucleotide diversity was found to be 0.02340 in the study population. The neutrality study found a significant level of nucleotide polymorphism, indicating a rise in low-frequency polymorphism, which might be attributed to global population growth of the parasite. Construction of phylogenetic comprising isolates from Kazakhstan, Finland, Turkiye, Canada, Germany, France, and China revealed that Pakistani isolates of H. taeniaeformis are distinct and formed a separate cluster. Further research utilising full-length multiple mitochondrial genes is required to understand the molecular epidemiology of H. taeniaeformis on the global scale. [ABSTRACT FROM AUTHOR]

Camel production in Pakistan is adversely affected by several pathogenic infections and insufficient veterinary facilities. Haemoparasitic diseases significantly affect health and productivity of camels causing a substantial financial burden to camel breeders and owners. The present study was designed for the identification and molecular detection of haemoparasites particularly piroplasms (Theileria spp. / Babesia spp.) infection in naturally infected local onehumped camels (Camelus dromedaries) in Punjab by using parasitological as well as molecular tools like polymerase chain reaction (PCR) followed by phylogenetic analysis. Blood samples (n=400) were collected from camels suspected for piroplasms infections in ten districts of Punjab and processed for blood smears and PCR targeting 18S rRNA gene. The findings revealed that Theileria is the most common parasite in camels of all study areas with overall prevalence of 12% and 13.5% by microscopic examination of GSBS and PCR, respectively. The phylogenetic analyses of the isolates on sequencing revealed that all analyzed isolates were closely related to Theileria annulata present in NCBI from several parts of the world. However, all samples tested for presence of Babesia spp. were found negative by microscopy and PCR. Chi square based risk factors analyses exhibited significant (P<0.05) association between gender, age, tick infestation, previous tick history and prevalence of Theileria. In conclusion, current study on haemoparasites is evident for first ever molecular identification of Theileria annulata infection in camels of Pakistan along with assessment of potential risk factors associated with disease. Recent outcomes are ascertaining it as a silent killer with damaging effects on immune system. [ABSTRACT FROM AUTHOR]

The organic and inorganic constituents present in cystic fluid play a crucial role in the physiology, metabolism and immune responses mediated by taeniids. We designed a comparative study on T. hydatigena and E. granulosus cysts based on biochemical profile. In the present study, we aspirated fluid from Taenia hydatigena (n=30) and Echinococcus granulosus (n=30) cysts recovered from slaughtered sheep and goats in the central abattoir of Faisalabad, Pakistan. The fluid obtained from each cyst was subjected to biochemical analysis to estimate the levels of liver enzymes, electrolytes and selected biomolecules (proteins, glucose, cholesterol, urea, and creatinine). Thereafter, the student's t-test was used to determine significant differences between constituents of the cystic fluids obtained from both type of cysts. The difference was considered statistically significant at P<0.05. The levels of total protein and globulin differed significantly (P<0.05) and were found to be higher in T. hydatigena cystic fluid. A non-significant difference was observed for albumin. Similarly, higher concentrations of electrolytes (sodium, chloride and magnesium) were observed in T. hydatigena cystic fluid with statistically significant difference (P<0.05). Concerning liver enzymes, high levels (p>0.05) of ALP and LDH were observed in T. hydatigena and E. granulosus cystic fluid had significantly higher (P<0.05) level of AST. The levels of cholesterol, urea and creatinine were higher in E. granulosus. In contrast, glucose was higher in T. hydatigena. A significant difference (P<0.05) was observed for these four biomolecules. Overall, these results suggest high variations of investigated biomolecules, liver enzymes, and electrolytes composition between cystic fluids of E. granulosus and T. hydatigena. Further studies are now warranted to set the standards for the differential diagnosis of cysticercosis and cystic echinococcosis based on cystic fluid contents. [ABSTRACT FROM AUTHOR]

Contagious caprine pleuropneumonia (CCPP) is an economically important and potentially fatal disease of small ruminants caused by Mycoplasma capricolum subsp. capripneumoniae (Mccp). We designed this cross-sectional study to investigate the seroepidemiology of CCPP in the small ruminants of Oman. For this purpose, we sampled a total of 4015 small ruminants (2119 goats and 1896 sheep) from 510 flocks belonging to different governorates of Oman. A commercial competitive enzyme-linked immune-sorbent assay (cELISA) was used to test the samples. Prevalence (%) along with 95% confidence intervals (CI) was calculated, and a univariable analysis was conducted to screen different risk factors. Furthermore, a binary logistic regression model was built at the animal and flocklevel (Table 3). In total, 147 (28.8%, CI 24.9, 33.0) flocks tested positive for the CCPP and the seroprevalence ranged from 10.0 to 53.8% in various governorates (p = 0.001). The prevalence was significantly (P<0.001) high in goats (28%, CI 23.8, 32.5) as compared to sheep (13.1%, CI 24.9, 33.0). At flock level; the open herds (OR 2.08, 1.33, 3.27), having a location in the coastal regions (OR 1.70, 1.14, 2.53) and flock size of more than 100 animals (OR 2.45, 1.54, 3.91) were the significant risk factors for CCPP in Oman. At the animal level; goats (OR 2.87, CI 2.17, 3.81), and small ruminants above the age of one year (OR 2.23, CI 1.38-3.59) were found more likely to acquire CCPP. We suggest that a control program based on the changes in the management system to minimize the risk factors and a possible mass vaccination should be devised to check CCPP in Oman. [ABSTRACT FROM AUTHOR]

Mosquitoes are highly active vectors capable of transmitting various pathogenic and infectious diseases to humans and animals. The present study was designed to identify the major species of mosquitoes and mosquito-borne flaviviruses (Saint Louis Encephalitis, West Nile and Dengue Viruses) prevalent in three districts of Punjab representing the three agro-geoclimatic zones viz; Multan, Chakwal and Jhang. The collected mosquitoes were stereoscopically identified to confirm the species and sex of the mosquitoes. Stereoscopic identification confirmed that female mosquitoes were 4334 out of 10675 (40.6%), 2242 out of 7296 (30%) and 2040 out of 6450 (31.6%) from districts Chakwal, Jhang and Multan, respectively. It was concluded that Culex species were present in abundance (73.2%) as compared to Aedes species (26.7%) in the selected study districts. The results of multiplex RT-PCR depicted that Dengue, West Nile, Japanese encephalitis and Saint Louis encephalitis viruses were prevalent in the mosquitoes of all three selected districts. However, the prevalence of mosquito-borne viruses insignificantly varied (P>0.05) among the three districts. This study has provided a better understanding of the prevailing mosquito species and mosquito-borne viruses in the study districts that can help to devise appropriate control measures. [ABSTRACT FROM AUTHOR]

Equine piroplasmosis is an endemic tick-borne disease of horses and other equids that is caused by Theileria equi and Babesia caballi. The study was aimed to investigate the seroprevalence of equine piroplasmosis during different seasons at district Sargodha located in central Punjab, Pakistan. For the detection of seropositive animals among sampled horses, the competitive enzyme-linked immunosorbent assay (cELISA) was performed against samples collected periodically during the four seasons of the year. On average basis, overall 36% of sampled horses were found seropositive with T. equi infection. Following the evaluation of seropositivity, subsequent estimation of oxidative stress biomarkers indicated that during winter season, total oxidant status (TOS) was found significantly (P≤0.05) increased while total antioxidant capacity (TAC), arylesterase and paraoxonase activity were found significantly decreased (P≤0.05) in seropositive horses as compared to healthy horses. Moreover, hematological parameters including red blood cells count, white blood cells count, MID and MID% were found significantly (P≤0.05) decreased while platelets, number of lymphocytes, number of granulocytes, and the percentage of lymphocytes were found significantly (P≤0.05) increased in seropositive horses as compared to seronegative horses in different seasons. In conclusion, we found that equine piroplasmosis markedly affect the oxidative stress biomarkers in seropositive horses primarily during the winter season as compared to all other seasons highlighting the seasonal impact of the disease. [ABSTRACT FROM AUTHOR]

Gastrointestinal nematodes (GINs) remain a major constraint associated with the profitable production of goats under grazing conditions. The present study was planned to determine the comparative susceptibility of Dera Din Panah (DDP) and Nachi breeds of goats towards artificial infection with Haemonchus (H.) contortus. To this end, a total of 24 goats of each breed were administered with third stage infective larvae of H. contortus through early and late infection protocols. The differences in faecal egg count (FEC), post necropsy worm count, rate of establishment of infection, packed cell volume (PCV), haemoglobin (Hb), eosinophils, total serum protein (TSP) and serum albumin (SA) were compared to check breed susceptibility to worms. Both breeds reflected significant (P<0.05) differences in aforementioned parameters at different time intervals post infection. However, Nachi breed showed a compromised response towards artificially infection with H. contortus as compared to DDP breed. Overall, higher FEC, higher number of adult worm recovery along with significant reduction in PCV and Hb depicted that Nachi is comparatively more susceptible to H. contortus infection as compared to DDP. In conclusion, difference in response towards H. contortus infection may formulate the base of selective breeding of resistant goat breed (DDP) in the area. [ABSTRACT FROM AUTHOR]

Brucellosis caused by Brucella melitensis is an important zoonotic disease in the world. Brucella melitensis is the causative organism of caprine and ovine brucellosis characterized by abortion, retained placenta, orchitis which may induce infertility, epididymitis and rarely, arthritis in small ruminants. However, these lesions were regarded as chronic while acute patho-morphological lesions had not been described in goats earlier. This study investigates the histopathological lesions in organs of native goats of Pakistan acutely infected by Brucella melitensis. Grossly, granulomatous lesions and edema were observed in the lungs of the infected animals. Other organs including liver, spleen, heart and spleen showed no gross lesions during necropsy. Severe histological lesions were observed in the lungs and uterus of acutely infected goats. The organism was detected by PCR from liver, lungs, spleen, uterus, sub mandibular lymph nodes and bronchial lymph nodes. The organism was not detected by PCR from the kidneys, ovaries, mammary glands, supra mammary lymph nodes, internal ileac lymph nodes and scapular lymph nodes. It was concluded that Br. melitensis acute infection did not lead to clinical presentation, although the organism was well distributed in various organs of goats. The lesions were generalized, not limited to reproductive organs. [ABSTRACT FROM AUTHOR]

Feline immunodeficiency virus (FIV) causes immunocompromising diseases in both domestic and wild cat species (Felis catus) around the world. Many characteristics of cats can affect the uptake of this virus including health status, age, gender, environment, lifestyle etc. The diagnosis of FIV can be confirmed by detecting antibodies in serum/plasma samples of infected cats. Despite its importance, the status of FIV infection is still poorly known in Pakistan. Therefore, the present study was devised to investigate the seroprevalence of FIV in Faisalabad, Pakistan. Blood samples of cats (n=90) were collected from outdoor facility of Department of Clinical Medicine and Surgery, University of Agriculture, Faisalabad and sera samples were analyzed for the presence of antibodies against FIV by ELISA Kit (Agrolabo S.p.A, Italy). The ELISA based results obtained by measuring the optical density of the substrate utilization showed the overall prevalence of FIV in cats as 28.9% in Faisalabad with a prevalence of 33.9% in male cats and 20.6% in female cats. The overall seropositivity for FIV was higher in adult cats (30.9%) than in young cats (22.7%). Risk factor association by Odd's ratio revealed that the disease is positively associated with adult age and male gender in cats. In conclusion, the factors of age (adult) and gender (male) were found to be the positively correlated (P=0.0164, OR=2.37, CI=0.74-7.55, RR=1.28, CI=0.94-1.73) with the presence of anti-FIV antibodies in the studied cats indicating the status of FIV infection in cats. [ABSTRACT FROM AUTHOR]

The present study was designed to investigate the ameliorative ability of locally available bentonite clay (BN) against aflatoxin B1 (AFB1) and ochratoxin A (OTA) toxicity in broiler chicks. In the first experiment the broiler chicks were kept on feeds contaminated with AFB1 (0.1, 0.2 and 0.6 mg/kg) alone and in combination with bentonite clay (3.7 and 7.5 g/kg). In the second experiment OTA (0.15, 0.3 and 1.0 mg/kg) alone and concurrently with bentonite clay (3.7, 7.5 and 15 g/kg) was fed to the broiler chicks. The dietary addition of AFB1 and OTA alone exerted deleterious effects on the performance, sero-biochemical and oxidative status of the broiler chicks. A concurrent dietary incorporation of BN (3.7 and 7.5 g/kg) ameliorated the adverse effects of 0.1 and 0.2 mg/kg AFB1 on body weight, FCR, serum ALT and urea concentration. Serum creatinine, total protein and albumin concentrations were mostly significantly lower in combination groups suggesting a partial or no amelioration. Dietary incorporation of BN at all levels only partially alleviated the deleterious effects of 0.15 and 0.3 mg/kg dietary OTA on performance, serobiochemical and oxidative stress, but amelioration was absent when the concentration of OTA in the feed was 1 mg/kg. Results suggested that the dietary incorporation of BN (3.7 and 7.5 g/kg) ameliorated the adverse effects of 0.1 and 0.2 mg/kg of AFB1 on various parameters. However, BN at all levels did not alleviate the deleterious effects of OTA. [ABSTRACT FROM AUTHOR]

Pertaining to reported high prevalence of brucellosis in Pakistan, district Layyah on account of having thick sheep population was selected for this study. A total of 384 sheep blood samples were collected randomly from different selected private herds in the district, and tested through Modified Rose Bengal (mRB) test and Indirect Enzyme Linked Immunosorbent Assay (iELISA) for the serological analysis against the Brucella antibodies. Positive samples from these two tests were further subjected to Competitive Enzyme Linked Immunosorbent Assay (cELISA). The individual based seroprevalence of brucellosis in sheep was found to be 7.0% by mRB. Herd based prevalence was 42.5%. The highest (P<0.05) seroprevalence (16.8%) was observed in Tehsil Layyah as compared to Tehsils Karor (2.3%) and Choubara (4.5%). Non-significant differences were recorded between breeds, age and sex groups and also for the animals with or without history of abortion. In case of indirect and competitive ELISA, no sheep serum sample was found to be positive. [ABSTRACT FROM AUTHOR]