Sequence analysis and biological characterization of virulent Avian avulavirus 1 isolated from asymptomatic migratory fowl.

Continuous monitoring and surveillance of avian avulaviruses (AAvVs) in water/migratory fowl is imperative to ascertain the evolutionary dynamics of these viruses. Here, we report genomic and amino acid characteristics of two AAvVs strains isolated from asymptomatic waterfowl (Anas carolinensis). Sequence characteristics including the presence of virulent motif (112RRQKR↓F117) and biological assessment confirmed the virulent nature of study isolates. Phylogenetic analysis of complete fusion (F) and hemagglutinin-neuraminidase (HN), and hyper-variable region of F gene revealed clustering of both strains within genotype VII and sub-genotype VIIi. The inferred residue analysis of complete F and HN genes revealed a number of substitutions in functionally and structurally important motif/s compared to reference strains of each genotype (I-XI). This study concludes an evolutionary nature of avian avulavaris 1 (AAvV-1), ascertaining continuous surveillance of migratory fowl to better elucidate their infection, epidemiology and subsequent impacts on commercial and backyard poultry. Keywords: virulent AAvV-1; migratory fowl; genetic characterization; evolutionary analysis; Pakistan.

Seroprevalence of Bluetongue Virus in small ruminants in Balochistan province, Pakistan.

Bluetongue (BT), caused by bluetongue virus (BTV), is a vector-borne disease of small ruminants that has the potential to spread across international borders. Despite large populations of susceptible animals and borders with BTV endemic countries, little is known of the disease burden and prevalent serotypes in the province of Balochistan in Pakistan. We conducted a cross-sectional study to determine seroconversion and prevalent serotypes in selected districts of the province using a competitive enzyme-linked immunosorbent assay (cELISA) and real-time polymerase chain reaction (RT-PCR). Sera (n = 876) were collected from clinically healthy sheep and goats originating from the districts of Quetta (n = 300), Mastung (n = 201), Killa Saifullah (n = 75) and Kech (n = 300). None of the study herds (n = 97) were seronegative for BTV, and at the individual level, the overall prevalence of BTV seroconversion was 47.26% (n = 414/876, 95% CI = 43.92%-50.63%). A higher percentage of goats (50.87%, 95% CI = 45.99%-55.73%) were seropositive for anti-VP7 immunoglobulins (IgG) than sheep (44.21%, 95% CI = 39.81%-48.70%). Odds ratios of seroconversion for goats were associated with breed type (χ2  = 16.84, p = .01), parity (χ2  = 23.66, p = .00) and presence of vector (χ2  = 2.63, p = .10), whereas for sheep, it was associated with breed type (χ2  = 13.80, p = .01) and parity (χ2  = 53.40, p = .00). Serotype 8 was the most prevalent (26.82%, 95% CI = 14.75%-43.21%) followed by an equal prevalence of serotypes 2 and 9 (7.31%, 95% CI = 1.91%-21.01%). To the best of our knowledge, this is the first study conducted in Balochistan province and the results indicate that there is a necessity to initiate intervention strategies to control BT disease burden not only in this region of Pakistan but also in adjacent areas of the neighbouring countries, Iran and Afghanistan.

Genetic characterization of canine parvovirus from dogs in Pakistan.

Canine parvoviruses (CPV) exist as antigenic variants with varying frequencies and genetic variabilities across the globe. Given the endemicity and high prevalence in Pakistan, we characterized the CPVs originating from dogs-population to elucidate viral diversity and evolution. Fecal samples from clinically diseased pups (n = 17) of different breeds and age (2-6 months) were processed for hemagglutination assay (HA), and later for partial amplification of VP2 gene sequence and amino acid analysis. A total of 11 samples (64.71%) were found positive both in hemagglutination and PCR assays. Phylogenetic and evolutionary analysis demonstrated higher genetic heterogeneity in studied strains and constituted seven clusters within the CPV-2a group, however, they shared high level of identity with Chinese strains. Further studies are necessary to elucidate genetic analysis and epidemiology of CPV variants across a wide geographical area of the country.

Molecular evidence shows that the liver fluke Fasciola gigantica is the predominant Fasciola species in ruminants from Pakistan.

Fascioliasis is an important disease affecting livestock, with great costs to producers worldwide. It has also become a serious issue for human populations in some endemic areas as an emerging zoonotic infection. There are two Fasciola species of liver fluke responsible for this disease, which occur worldwide, Fasciola hepatica and Fasciola gigantica. Identifying these two species on the basis of adult or egg morphology requires specialist knowledge due to the similarity of characters, and may misidentify putative intermediate or hybrid forms. In this study we sequenced the internal transcribed spacer 2 (ITS-2) rDNA of liver flukes collected from multiple species of hosts from seven localities in the Punjab and Baluchistan provinces of Pakistan, to determine the distribution of these two species. All 46 flukes processed in this study, collected from seven sites, showed the rDNA ITS-2 genotype corresponding to F. gigantica, contradicting previous reports, based on adult and egg morphology, that both species are present in Pakistan, with F. hepatica being the more common.

Effect of supplementation of prebiotic mannan-oligosaccharides and probiotic mixture on growth performance of broilers subjected to chronic heat stress.

The present study was aimed at elucidating the effects of supplementing mannan-oligosaccharides (MOS) and probiotic mixture (PM) on growth performance, intestinal histology, and corticosterone concentrations in broilers kept under chronic heat stress (HS). Four hundred fifty 1-d-old chicks were divided into 5 treatment groups and fed a corn-soybean diet ad-libitum. The temperature control (CONT) group was held at the normal ambient temperature. Heat stress broilers were held at 35 ± 2°C from d 1 until the termination of the study at d 42. Heat stress groups consisted of HS-CONT fed the basal diet; HS-MOS fed the basal diet containing 0.5% MOS; HS-PM fed the basal diet containing 0.1% PM; and HS-SYN (synbiotic) fed 0.5% MOS and 0.1% PM in the basal diet. Broilers were examined at d 21 and 42 for BW gain, feed consumption, feed conversion ratio (FCR), serum corticosterone concentrations, and ileal microarchitecture. The results revealed that the CONT group had higher (P < 0.01) feed consumption, BW gain, and lower FCR on d 21 and 42, compared with the HS-CONT group. Among supplemented groups, the HS-MOS had higher (P < 0.05) BW gain and lower FCR compared with the HS-CONT group. On d 21 and 42, the HS-CONT group had higher (P < 0.05) serum corticosterone concentrations compared with the CONT and supplemented groups. The CONT group had higher (P < 0.05) villus height, width, surface area, and crypt depth compared with the HS-CONT group. On d 21, the HS-PM had higher (P < 0.05) villus width and surface area compared with HS-CONT group. On d 42, the HS-SYN had higher (P < 0.05) villus width and crypt depth compared with the HS-CONT group. These results showed that chronic HS reduces broiler production performance, intestinal microarchitecture, and increases adrenal hormone concentrations. Also, supplementation of the MOS prebiotic and the PM can partially lessen these changes.