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Blood and fecal samples of chukar partridge (Alectoris chukar), albino pheasant (Phasianus colchicus), silver pheasant (Lophura nycthemera), rose-ringed parakeet (Psittacula krameri) and turkeys (Meleagris gallopavo) were analyzed to check parasitic prevalence. To record parasites these five avian species were placed kept in separate cages at Avian Conservation and Research Center, Department of Wildlife an Ecology, University of Veterinary and Animal Sciences, Lahore, Pakistan. 100 fecal and 100 blood samples for each bird species were inspected to analyze internal parasites. During present study, 17 species of endoparasites 14 from fecal samples and three from blood were examined. Two species of ectoparasites i.e. mite Dermanyssus gallinae 42% and fowl ticks Args persicus 41%were studied. Blood parasites included Plasmodium juxtanucleare 50%, Leucoctoyzoon simond having parasitic prevalence 40%, and Aegyptinella pullorum having parasitic prevalence of 40%. Parasitic species recorded from fecal samples included 6 species of nematodes viz. Allodpa suctoria 2%. Syngamus trachea with parasitic prevalence of 60%, Capillaria annulata 37.5%, Ascardia galli 24%, Capillaria anatis 40% and Heterakis gallinarum 28.3%. Similarly, two species of trematodes viz. Prosthogonimus ovatus having parasitic prevalence of 50% and Prosthogonimus macrorchis 21% were also documented from fecal avian samples . Single cestode species Raillietina echinobothrida having parasitic prevalence of 72% and 3 protozoan species i.e. Eimeria maxima having parasitic prevalence of 21%, Giardia lamblia 41% and Histomonas meleagridis 18% were documented during corpological analysis. In our recommendation, proper sanitation, medication and vaccination of bird’s enclousres are suggested to avoid parasites. RESUMO Amostras de sangue e fezes de perdiz chukar (Alectoris chukar), faisão-albino (Phasianus colchicus), faisão-prateado (Lophura nycthemera), periquito-de-rosa (Psittacula krameri) e perus (Meleagris gallopavo) foram analisadas para verificar a prevalência de parasitas. Para registrar os parasitas, essas cinco espécies de aves foram colocadas em gaiolas separadas no Centro de Conservação e Pesquisa de Aves, Departamento de Vida Selvagem e Ecologia, Universidade de Veterinária e Ciências Animais, Lahore, Paquistão. Cem amostras fecais e 100 amostras de sangue para cada espécie de ave foram inspecionadas para analisar os parasitas internos. Durante o presente estudo, foram examinadas 17 espécies de endoparasitas, 14 de amostras fecais e 3 de sangue. Foram estudadas duas espécies de ectoparasitas, ou seja, o ácaro Dermanyssus gallinae 42% e o carrapato aviário Args persicus 41%. Os parasitas sanguíneos incluíram Plasmodium juxtanucleare 50%, Leucoctoyzoon simond com prevalência parasitária de 40% e Aegyptinella pullorum com prevalência parasitária de 40%. As espécies parasitas registradas em amostras fecais incluíram 6 espécies de nematoides viz. Allodpa suctoria 2%, Syngamus traqueia com prevalência parasitária de 60%, Capillaria annulata 37,5%, Ascardia galli 24%, Capillaria anatis 40% e Heterakis gallinarum 28,3%. Da mesma forma, duas espécies de trematódeos viz. Prosthogonimus ovatus com prevalência parasitária de 50% e Prosthogonimus macrorchis 21% também foram documentados em amostras fecais de aves. Espécies de cestoide único Raillietina echinobothrida com prevalência parasitária de 72% e 3 espécies de protozoários, isto é, Eimeria maxima com prevalência parasitária de 21%, Giardia lamblia 41% e Histomonas meleagridis 18% foram documentadas durante a análise corpológica. Em nossa recomendação, o saneamento adequado, medicação e vacinação de invólucros de pássaros são sugeridos para evitar parasitas.

In this study, we reported the isolation, identification, and molecular characteristics of nine BVDV strains that were isolated from the serum of persistently infected cattle. The new strains were designated as BVDV TJ2101, TJ2102, TJ2103, TJ2104, TJ2105, TJ2106, TJ2107, TJ2108 and TJ2109. The TJ2102 and TJ2104 strains were found to be cytopathic BVDV, and the other strains were non-cytopathic BVDV. An alignment and phylogenetic analysis showed that the new isolates share 92.2–96.3% homology with the CP7 strain and, thus, were classified as the BVDV-1b subgenotype. A recombination analysis of the genome sequences showed that the new strains could be recombined by the major parent BVDV-1a NADL strain and the minor parent BVDV-1m SD-15 strain. Some genome variations or unique amino acid mutations were found in 5′-UTR, E0 and E2 of these new isolates. In addition, a potential linear B cell epitopes prediction showed that the potential linear B cell epitope at positions 56–61 is highly variable in BVDV-1b. In conclusion, the present study has identified nine strains of BVDV from persistently infected cattle in China. Further studies on the virulence and pathogenesis of these new strains are recommended.

Buffalo animals are slaughtered at their early age and carcasses are chilled rapidly which deteriorates its meat quality and decreases the consumer likeliness of buffalo meat. This study investigated the appropriate methods to prevent the quality deterioration of buffalo meat during chilling. Twenty four 18-mon-old buffalo bulls were slaughtered, electrically stimulated and suspended either by hip or achilles tendon. After 24 h postmortem, meat quality characteristics were recorded. Results showed that electrical stimulation (ES) led to rapid decline of carcass pH compared to non-ES method (p

The potential spillover of foot and mouth disease (FMD) virus at the wildlife-livestock interface is mainly responsible for the outbreaks in captive wild ungulates. The current study was planned to investigate an FMD outbreak in the wild ungulate species in the Jallo Wildlife Park and breeding facility, Lahore, Pakistan from Mar 2021- Jun 2021. The disease was confirmed based on ELISA through the detection of antibodies against the non-structural protein of FMD virus and typical clinical signs of oral and feet lesions, and even partial or complete hoof shedding in severe cases. To investigate the possible cause of FMD spillover and its pattern, a series of interviews were conducted with wildlife practitioners, animal handlers, and the local veterinarians in villages adjacent to the park. The data revealed neither vaccination nor any FMD outbreak in the last 10 years. The epidemic curve and time-series analysis showed a mixed outbreak characterized by the introduction of disease and then propagative spread from infected to susceptible animals. After the first reports of clinical signs in Blackbuck, Urial and Mouflon sheep were infected while in the end, the cases were reported from the enclosures of Sambar deer and Spotted deer. The morbidity rate among all the wild ungulate species was 92% and the mean mortality rate was 27%. The study concluded that possible sources for primary disease incidence might be animal handlers having FMD infected animals in their houses and transport vehicles coming from adjacent disease-affected villages. For the disease spread between enclosures, another possibility of wind-borne FMD virus spread could also be considered. This is the first report regarding the FMD outbreak and its spillover pattern in captive wild ungulates in Pakistan. These findings will be helpful in understanding the importance of wildlife livestock interface in FMD transmission and to design effective strategies to control the disease.

Efforts to identify biomarkers for neuroblastoma (NB) have been ongoing, but no definite biomarker has been identified in peripheral blood. We proposed the use of plasma exosomal miRNAs as biomarkers of unfavorable NB patient outcomes. Exosomal miRNAs isolated from 31 plasma and 37 tissue samples, many from the same NB patients, were sequenced using a next-generation sequencing instrument. We analyzed the correlation between miRNA expression levels in plasma and tissue samples with International Neuroblastoma Risk Group staging system (INRGSS) outcome and MYCN status. We chose differentially expressed miRNAs with similar expression patterns in plasma and tissue samples in each of the three analysis groups and combined those miRNAs to find the optimal combination with the potential to be considered as a biomarker. MicroRNA-92a-3p was found to be significantly upregulated in deceased patients (p = 0.017), miR-375 was upregulated in INRGSS stage M patients (p = 0.002), and plasma miR-92a-3p and miR-99a-5 levels were upregulated in patients with MYCN amplification (p = 0.007 and 0.006). The combination of miR-92a-3p, miR-375, and miR-99a-5p levels was shown to be a statistically significant predictor of NB patient outcomes (AUC = 0.726, p = 0.001, 95% CI = 0.612–0.841, sensitivity = 77%, specificity = 56.7%). Thus, the combination of miR-92a3p, miR-375, and miR-99a-5p may potentially be used as a biomarker for unfavorable NB patient outcomes. However, further validation is required in a larger number of NB patients., This research was partially supported by a Grant-in-Aid for Scientific Research (A) (No.19H0105600) from the Ministry of Education, Culture, Sports, Science and Technology.

Myostatin is a protein translated by the the MSTN gene (also known as GDF8), is responsible for limiting muscle growth and strength. In thoroughbred horse (Equus Caballus), limited studies have been designed to examine the variants in the coding region of MSTN gene. However, no data is available regarding the single nucleotide polymorphisms (SNPs) of the MSTN about racing performance in thoroughbred horses of Pakistan. In this study blood samples of fifteen Pakistani thoroughbred horses were collected from Race Club Lahore and immediately transferred into the ice box. The DNA was extracted by using phenol-chloroform method. Primers were designed for the amplification of all exons of the MSTN gene. The amplified PCR products were precipitated and sequenced for the identification of SNPs. SNPs were identified by visualizing the peaks of sequenced data by using Chromas Software. Phylogenetic analysis of MSTN gene in Pak-thoroughbred with racing species and some breeds of horses like Marwari Indian breed, Sindhi breed, Kathlawari breed, Italian breed and Chines breed was done separately by using MEGA 6 software. The analysis of identified SNPs were carried out by software SNPator. The sequenced data with altered protein was published in GenBank with accession number MN604194. Results have shown a total of 3 single nucleotide polymorphisms through Blast with reference sequences. Two SNPs were found in exon 2 at position of 2406 (C/T) and 2408 (C/T) respectively. One SNP (T/C) was detected in exon 3 at the position of 4661. In conclusion, Pak-thoroughbred horse population has 3 polymorphisms in their coding region which can be used as a biological marker for athletic abilities in Pak- thoroughbred.

The medicinal attributes of honey appears to overshadow its importance as a functional food. Consequently, several literatures are rife with ancient uses of honey as complementary and alternative medicine, with relevance to modern day health care, supported by evidence-based clinical data, with little attention given to honey’s nutritional functions. The moisture contents of honey extracted from University of Veterinary and Animal Sciences, Lahore honey bee farm was 12.19% while that of natural source was 9.03 ± 1.63%. Similarly, ash and protein contents of farmed honey recorded were 0.37% and 5.22%, respectively. Whereas ash and protein contents of natural honey were 1.70 ± 1.98% and 6.10 ± 0.79%. Likewise fat, dietary fiber and carbohydrates contents of farmed source documented were 0.14%, 1.99% and 62.26% respectively. Although fat, dietary fiber and carbohydrates contents of honey taken from natural resource were 0.54 ± 0.28%, 2.76 ± 1.07% and 55.32 ± 2.91% respectively. Glucose and fructose contents of honey taken out from honeybee farm were 27% and 34% but natural source were 22.50 ± 2.12% and 28.50 ± 3.54%. Glucose and fructose contents of honey taken out from honeybee farm were 27% and 34% but natural source were 22.50 ± 2.12% and 28.50 ± 3.54%. Similarly, sucrose and maltose contents of farmed honey were 2.5% and 12% while in natural honey were 1.35 ± 0.49% and 8.00 ± 1.41% respectively. The present study indicates that such as moisture, carbohydrates, sucrose and maltose contents were higher farmed honey as compared to the natural honey. In our recommendation natural honey is better than farmed honey. Resumo Os atributos medicinais do mel parecem ofuscar sua importância como alimento funcional. Consequentemente, várias literaturas estão repletas de usos antigos do mel como medicina complementar e alternativa, com relevância para os cuidados de saúde modernos, apoiados por dados clínicos baseados em evidências, com pouca atenção dada às funções nutricionais do mel. O teor de umidade do mel extraído da Universidade de Veterinária e Ciências Animais, fazenda de abelhas de Lahore, foi de 12,19%, enquanto o de fonte natural foi de 9,03 ± 1,63%. Da mesma forma, os teores de cinzas e proteínas do mel cultivado foram de 0,37% e 5,22%, respectivamente. Já os teores de cinzas e proteínas do mel natural foram de 1,70 ± 1,98% e 6,10 ± 0,79%. Da mesma forma, os teores de gordura, fibra dietética e carboidratos de origem cultivada documentados foram de 0,14%, 1,99% e 62,26%, respectivamente. Embora os teores de gordura, fibra alimentar e carboidratos do mel retirado dos recursos naturais fossem de 0,54 ± 0,28%, 2,76 ± 1,07% e 55,32 ± 2,91%, respectivamente. Os conteúdos de glicose e frutose do mel retirado da fazenda de abelhas foram de 27% e 34%, mas a fonte natural foi de 22,50 ± 2,12% e 28,50 ± 3,54%. Os conteúdos de glicose e frutose do mel retirado da fazenda de abelhas foram de 27% e 34%, mas a fonte natural foi de 22,50 ± 2,12% e 28,50 ± 3,54%. Da mesma forma, os teores de sacarose e maltose no mel cultivado foram de 2,5% e 12%, enquanto no mel natural foram de 1,35 ± 0,49% e 8,00 ± 1,41%, respectivamente. O presente estudo indica que os teores de umidade, carboidratos, sacarose e maltose foram maiores no mel cultivado em comparação ao mel natural. Em nossa recomendação, o mel natural é melhor que o mel de cultivo.

Newcastle disease virus, a prototype avian paramyxovirus serotype 1 (APMV-1), causes economically devastating disease in avian species around the world. Newcastle disease is enzootic in Pakistan and recurrent outbreaks are frequent in multiple avian species even after continuous and extensive use of vaccines. A number of APMV-1 and pigeon paramyxovirus serotype 1 (PPMV-1) strains have been isolated and genetically characterized in recent years. However, the impact of recently characterized wild bird-origin APMVs in domestic poultry, and the potency of routinely used vaccines against these novel and genetically diverse viruses remain unknown. Here, we applied next-generation sequencing for unbiased complete genome characterization of APMV-1 and PPMV-1 strains isolated from clinically diseased peacocks (Pavocristatus) and pigeons (Columbalivia), respectively. Global phylodynamics and evolutionary analysis demonstrates Pigeon/MZS-UVAS-Pak/2014 is clustered into lineage 4 (or genotype VI) and Peacock/MZS-UVAS-Pak/2014 into lineage 5 (or genotype VII). The genomes of both isolates encoded for polybasic residues (112RRQKR↓F117) at the fusion protein cleavage motif along with a number of important substitutions in the surface glycoproteins compared with the vaccine strains. Clinicopathological and immunological investigations in domesticated chickens indicate that these isolates can potentially transmit between tested avian species, can cause systemic infections, and can induce antibodies that are unable to prevent virus shedding. Collectively, the data from these genomic and biological assessments highlight the potential of wild birds in transmitting APMVs to domesticated chickens. The study also demonstrates that the current vaccine regimens are incapable of providing complete protection against wild bird-origin APMVs and PPMVs.

Coxiella burnetii causes query (Q) fever, an important zoonotic disease with worldwide significance. The role of environment in the ecology of C. burnetti, and its influence on seroconversion in animals has not been elucidated in Pakistan. We carried out a cross-sectional study in Punjab province to (1) determine the prevalence and distribution of C. burnetii in soil using an ISIIII gene-based real time-polymerase chain reaction (RT-PCR) assay, (2) analyze association between the occurrence of C. burnetii in soil and its predictors i.e. soil characteristics (macro- and micro-nutrients) and several likely risk factors including the seroconversion in small ruminants at places where its genome had or had not been detected, and (3) predict homology and genetic diversity of the identified strains using sequences originated from different hosts worldwide. A total of 2425 soil samples from nine districts of Punjab province were processed. C. burnetii DNA was detected in 47 samples (1.94%, 95% CI: ±0.55) originating from 35 villages of studied districts (7.22%, 95% CI: ±2.30). The highest prevalence was found in Attock (7.11%, 95% CI: ±3.36), followed by Lahore (4.83%, 95% CI: ±3.49), Sahiwal (4.70%, 95% CI: ±2.6), Dera Ghazi Khan (2.33%, 95% CI: ±2.02), Faisalabad (1.35%, 95% CI: ±1.18) and Sheikhupura (0.68%, 95% CI: ±0.94). The odds of detecting bacterial DNA in soil was increased with a unit increase in organic matter [2.511 (95% CI: 1.453-4.340), p=0.001] and sodium [1.013 (95% CI: 1.005-1.022), p=0.001], whereas, calcium [0.984 (95% CI: 0.975-0.994), p=0.002] and potassium [0.994 (95% CI: 0.990-0.999), p=0.011] had protective effect where a unit increase in each analyte decreased odds for its occurrence by 1.0% approximately. Likewise, for categorical variables (risk factors), the odds of detecting C. burnetii were higher at locations >500m away from a main road [1.95 (95% CI: 1.06-3.78), p=0.04]. The enzyme-linked immunosorbent assay (ELISA) revealed an increased prevalence of antibodies in sheep (17.9%, 95% CI: ±5.54) compared with goats (16.4%, 95% CI: ±4.34). When determining the association between soil DNA and C. burnetii antibodies in small ruminants, the odds of detecting these antibodies were significant in sheep at the livestock barns [2.81 (95% CI: 1.20-7.37), p=0.02]. The IS1111 gene-based sequence analysis revealed a clustering of the DNA into two distinct groups with much genetic divergence (0.76-68.70%): the first group that contained sequences from Lahore district clustered with human and buffalo origin isolates, whereas the second group that contained the sequences from the remaining study districts clustered with goat-, rodent- and human-origin isolates. This study provides the first evidence of the presence of C. burnetii in the environment in Punjab province, Pakistan. Future studies are needed to ascertain the bacteria's molecular epidemiology over a wide geographical area, type the isolates, and evaluates the potential risks to human populations, particularly farmers and veterinarians.

Association of soil chemistry and other risk factors with soil borne Burkholderia mallei (B. mallei) DNA in eight districts (Sheikhupura, Gujranwala, Faisalabad, Sargodha, Chakwal, Attock, Sahiwal, Dera Ghazi Khan) of Punjab province, Pakistan, was studied. A total of 22 soil samples (n=11, each from soil positive and negative for B. mallei DNA) were processed for chemical analysis from B. mallei positive districts (Sheikhupura and Chakwal). The relationship between soil composition, absence or presence of the pathogen was ascertained. In soil samples of Chakwal district, DNA of B. mallei was found to be highly associated with 0.80 to 39.20% range of moisture contents (p=0.008) and 1.74 to 21.75 mg/Kg of P (p=0.050). The association in Sheikhupura district was with sodium (1.90 to 133.59 mg/Kg; p=0.018) and moisture (0.80 to 39.20%; p=0.026). The odds of detecting DNA of B. mallei was 1.4, 6.8, 5.0, 2.8 and 10.6 higher when soil sample sites were 500 meters from vehicular traffic roads, > 01 km from animal markets, > 100 meters from canal, > 1000 animals and > 300 houses/village. Soilborne B. mallei DNA is more likely to be detected in areas closer to roads having vehicular traffic along with interstate routes and soil containing low levels of moisture.

Influenza A viruses infect various mammals like human, horse, pig and birds as well. A total of 16 hemagglutinin (HA) and 9 neuraminidase (NA) subtypes have been identified. Most of the combinations are found in birds and relatively few have been isolated from mammals. Although there is no report of human to human transmission till to date, several cases of H5N1, H7N7 and H9N2 identified in humans since 1997 raised serious concern for health and veterinary profession. This review paper will focus H9N2 avian influenza virus (AIV) with special emphasis on zoonosis. The virus H9N2 though not highly pathogenic like H5N1 but can be virulent through antigenic drift and shift.

Abortion is common among dairy herds in Pakistan. However, except for Brucella abortus, little is known about other prevalent abortifacients. Therefore, a sero-epidemiological study was conducted in a dairy herd with a history of abortion located in Lahore, Pakistan. Blood samples (3–5 mL) were taken from each animal (cows, n = 43; buffaloes, n = 47) in the herd. Seropositivity to infectious bovine rhinotracheitis (IBR), B. abortus, blue tongue virus (BTV), bovine viral diarrhea virus (BVDV) and Toxoplasma gondii was determined using commercially available kits. Among cows and buffaloes, seropositivity was most frequent for IBR (69.8 and 70.3%, P>0.05), followed by B. abortus (32.6 and 42.6%, P>0.05), BVDV (9.3 and 6.4%, P>0.05) and BTV (4.7 and 6.4%, P>0.05), whereas, all the animals were seronegative to T. gondii. With respect to age, there was a significant difference (P

WOS: 000368558800001, Influenza virus H9 was standardized for hemagglutination assay using different factors such as red blood cell (RBC) types, concentrations, diluent types, and storage times. Avian influenza virus H9 was grown in embryonated chicken eggs and confirmed by spot agglutination. A significant (P < 0.05) difference with the highest mean titer (9.00 perpendicular to 0.00) was observed using RBCs from different species. Nonsignificant differences (P > 0.05) were found between human blood type O, chicken, and dog RBCs, as well as among rabbit, pigeon, sheep, and parrot. The highest titer (9.00 +/- 0.00) with a nonsignificant difference was found using virus stored at 4 degrees C and -20 degrees C while 37 degrees C showed the lowest significant mean hemagglutinin (HA) titer (11.08 +/- 188.21). Nonsignificant differences were observed in HA titers against H9 virus stored for 3, 4, 5, and 6 months. Nonsignificant differences were found between the use of normal saline and 0.5% peptone water with the lowest HA titers of 7.83 +/- 0.40 and 8.00 +/- 0.00, respectively, while the highest HA titer (9.00 +/- 0.00) with nonsignificant difference was observed by using HA-HI buffer and phosphate buffer saline as diluents. RBCs with 0.5% and 1% concentrations showed nonsignificant difference in HA titer but significant difference with 0.1% RBCs.

The effects of time elapsed after milking and lactation stage were assessed on the lactoperoxidase (LP) enzyme activity and concentrations of thiocyanate in Sahiwal cattle (N = 15) and Nili-Ravi buffaloes (N = 15). The LP activity and thiocyanate concentrations decreased as the time elapsed after milking (P

A multidisciplinary, collaborative project was conducted to determine the prevalence and distribution of soil-borne zoonotic pathogens in Lahore district of Pakistan and ascertain its Public Health Significance. Using a grid-based sampling strategy, soil samples (n = 145) were collected from villages (n = 29, 5 samples/village) and examined for Bacillus anthracis, Burkholderia mallei/pseudomallei, Coxiella burnetii, Francisella tularensis, and Yersinia pestis using real time PCR assays. Chemical analysis of soil samples was also performed on these samples. The relationship between soil composition and absence or presence of the pathogen, and seven risk factors was evaluated. DNA of B. anthracis (CapB), B. mallei/pseudomallei (chromosomal gene), C. burnetii (IS1111, transposase gene), and F. tularensis (lipoprotein/outer membrane protein) was detected in 9.6, 1.4, 4.8, and 13.1% of soil samples, respectively. None of the samples were positive for protective antigen plasmid (PA) of B. anthracis and Y. pestis (plasminogen activating factor, pPla gene). The prevalence of B. anthracis (CapB) was found to be associated with organic matter, magnesium (Mg), copper (Cu), chromium (Cr), manganese (Mn), cobalt (Co), cadmium (Cd), sodium (Na), ferrous (Fe), calcium (Ca), and potassium (K). Phosphorous (P) was found to be associated with prevalence of F. tularensis while it were Mg, Co, Na, Fe, Ca, and K for C. burnetii. The odds of detecting DNA of F. tularensis were 2.7, 4.1, and 2.7 higher when soil sample sites were >1 km from animal markets, >500 m from vehicular traffic roads and animal density of < 1000 animals, respectively. While the odds of detecting DNA of C. burnetii was 32, 11.8, and 5.9 higher when soil sample sites were >500 m from vehicular traffic roads, presence of ground cover and animal density of < 1000 animals, respectively. In conclusion, the distribution pattern of the soil-borne pathogens in and around the areas of Lahore district puts both human and animal populations at a high risk of exposure. Further studies are needed to explore the genetic nature and molecular diversity of prevailing pathogens together with their seroconversion in animals and humans.

The protozoan Neospora caninum and the bacterium Brucella abortus are well-recognized causes of abortion in dairy cattle. Serum samples (n = 240) from aborting (n = 141) and at-risk (n = 99) animals from 5 herds with high abortion rates in Punjab Province, Pakistan, were tested for antibodies to N. caninum using monoclonal antibody-based ELISA and for antibodies to B. abortus using the serum agglutination test. Antibodies to N. caninum and B. abortus were detected in 105 (43.8%) and 135 (56.3%) cattle, respectively. Prevalences of antibodies to N. caninum and B. abortus were higher in aborting cows (46.8% and 76.6%, P0.05) than in animals at risk (39.4% and 27.3%, P0.05). Sixty-six animals (27.5%) were seropositive to both N. caninum and B. abortus , and results showed no significant difference (P0.05) with respect to geographical district, breed, and age. This is the first report of N. caninum infection among dairy cattle herds in Pakistan.

We report the complete genome sequence of avian paramyxovirus 1 (APMV-1) isolated from an acute and highly contagious outbreak in pheasants ( Pucrasia macrolopha ) in Lahore, Pakistan. Biological and serological characterization showed a velogenic strain of APMV-1, which was further confirmed by the sequence analysis of the cleavage site in the fusion protein. Complete genome sequencing and phylogenetic analysis indicated that this isolate belonged to genotype VII, specifically to subgenotype VIIa, and clustered closely with isolates characterized from Indonesia. Notably, the isolate showed significant differences from previously characterized APMV-1 from Pakistani commercial and rural chicken.

Background: Newcastle disease (ND) is one of the most deadly diseases of poultry around the globe. The disease is endemic in Pakistan and recurrent outbreaks are being reported regularly in wild captive, rural and commercial poultry flocks. Though, efforts have been made to characterize the causative agent in some of parts of the country, the genetic nature of strains circulating throughout Pakistan is currently lacking. Material and methods: To ascertain the genetics of NDV, 452 blood samples were collected from 113 flocks, originating from all the provinces of Pakistan, showing high mortality (30 – 80%). The samples represented domesticated poultry (broiler, layer and rural) as well as wild captive birds (pigeons, turkeys, pheasants and peacock). Samples were screened with real-time PCR for both matrix and fusion genes (1792 bp), positive samples were subjected to amplification of full fusion gene and subsequent sequencing and phylogenetic analysis. Results: The deduced amino acid sequence of the fusion protein cleavage site indicated the presence of motif (112 RK/RQRR↓F117) typical for velogenic strains of NDV. Phylogenetic analysis of hypervariable region of the fusion gene indicated that all the isolates belong to lineage 5 of NDV except isolates collected from Khyber Pakhtunkhwa (KPK) province. A higher resolution of the phylogenetic analysis of lineage 5 showed the distribution of Pakistani NDV strains to 5b. However, the isolates from KPK belonged to lineage 4c; the first report of such lineage from this province. Conclusions: Taken together, data indicated the prevalence of multiple lineages of NDV in different poultry population including wild captive birds. Such understanding is crucial to underpin the nature of circulating strains of NDV, their potential for interspecies transmission and disease diagnosis and control strategies. Peer Reviewed publishedVersion