Introduction: Leptospirosis affects a wide range of mammals, humans, and even a few poikilothermic animal species. In Pakistan, serological studies of equine leptospirosis have reported a prevalence of over 40%, but no study has ever been conducted towards molecular detection of Leptospira in horses. Material and Methods: Blood samples from 128 horses were screened using ELISA and 41 positive samples were examined for the presence of leptospiral DNA using specific primers for 16S rRNA gene. Results: Out of 41 tested samples, 20 samples were found to be PCR-positive, revealing a fragment of 306 bp after gel electrophoresis. Sequencing and phylogenetic analysis of positive samples revealed circulation of pathogenic Leptospira spp. in Pakistani horses. No evidence of circulation of intermediate species was found in this study. Conclusion: This study reports the first molecular evidence of equine leptospirosis in Pakistan and lays ground for further research in this area. It also confirms the efficiency of 16S rRNA for the diagnosis of equine leptospirosis.

Introduction: Leptospirosis affects a wide range of mammals, humans, and even a few poikilothermic animal species. In Pakistan, serological studies of equine leptospirosis have reported a prevalence of over 40%, but no study has ever been conducted towards molecular detection of Leptospira in horses. Material and Methods: Blood samples from 128 horses were screened using ELISA and 41 positive samples were examined for the presence of leptospiral DNA using specific primers for 16S rRNA gene. Results: Out of 41 tested samples, 20 samples were found to be PCR-positive, revealing a fragment of 306 bp after gel electrophoresis. Sequencing and phylogenetic analysis of positive samples revealed circulation of pathogenic Leptospira spp. in Pakistani horses. No evidence of circulation of intermediate species was found in this study. Conclusion: This study reports the first molecular evidence of equine leptospirosis in Pakistan and lays ground for further research in this area. It also confirms the efficiency of 16S rRNA for the diagnosis of equine leptospirosis.

OBJECTIVE To develop 2 rapid loop-mediated isothermal amplification (LAMP) assays for detection of Pasteurella multocida DNA (Pm-LAMP assay) and P multocida DNA from strains associated with hemorrhagic septicemia (HS) in cattle and buffalo (HS-LAMP assay). SAMPLE Solutions containing 16 P multocida strains and 9 other bacterial species at various concentrations. PROCEDURES Optimal conditions were determined for running the Pm-LAMP and HS-LAMP assays. The assays were then used to detect DNA of the test organisms. Results of LAMP assays were validated against conventional PCR assays designed for specific detection of P multocida and the B:2 serotype of HS-associated strains. RESULTS Following incubation of sample reaction mixtures for 27 minutes, specificity and sensitivity of the HS-LAMP assay at template DNA amounts as low as 5 pg were 93% and 97%, respectively. When duplicates of each sample were incubated for 28 minutes (a positive result defined as positive results for both reactions of a given sample), specificity and sensitivity of the HS-LAMP assay in the same conditions increased to 100%. The best specificity and sensitivity of Pm-LAMP single (93% and 91%) and duplicate (97% and 98%) reactions at template DNA amounts as low as 10 pg were achieved at 33 and 34 minutes, respectively. CONCLUSIONS AND CLINICAL RELEVANCE These preliminary findings suggested the developed HS-LAMP assay had high sensitivity and specificity for detection of HS-associated P multocida. Additional research is needed to determine the accuracy of the assay for use on clinical specimens obtained in HS-endemic countries such as Pakistan and Thailand.

A study was conducted to determine the effect of different nutrients i.e. metabolizable energy (ME) and crude protein (CP) content of ration on the egg production and egg weight of rural layers. For the purpose six different experimental rations containing 2700, 2900 kcal/kg ME and 14, 16, 18 percent CP were offered to the birds. The results indicated that egg number increased with proportional adversely affected as the CP of the ration was increased weight of the layers was proportionally increased with ration at constant ME level, while increase in ME level egg weight. increase in ME, while it was at constant ME level. The egg increase in CP content of the did not significantly affect the egg weight.

This study was conducted to ascertain anatomical variation in different body parts of Desi, Fayoumi, Cross (Rhode Island Red x Fayoumi) and Nick Chick laying hens. A total 16 laying hens of average body weight were selected using random numbers out of 2600 laying hens, slaughtered and eviscerated. It was observed that proportion of neck, ribs, breast, back, wings, thigh and legs out of aggregate weight was highest (51.74 plus minus 2.85) in Fayoumi hens. The proportion of liver, gizzard, heart and spleen combined weight was highest (6.05 plus minus 1.81) in desi hens. The proportion of non edible offals including trachea, lungs, kidneys, blood, feathers, head, crop, proventriculus, intestines, shanks, gizzard waste, skin, tail, testes, ova, oviduct and fat was estimated to be highest i.e. 46.60 plus minus 1.16 in Nick Chick laying hens. The anatomical variation in Desi and Fayoumi chicks was attributable to scavenging habit of these birds. The high proportion of edible offal in commercial Nick Chick hens may be ascribable to higher productive and reproductive traits.

This study was carried out for investigation of an outbreak in Karachi. A disease outbreak in poultry was reported during April, 2000 in Karachi. The main symptoms included respiratory distress, sneezing and gasping. Autopsy of dead birds showed tracheitis, air-sacculitis and involvement of lungs. The morbid material was collected for processing in the laboratory. During the investigation for isolation of pathogenic bacteria or virus, Ornithobacterium rhinotracheale was isolated on 5% sheep blood agar plates in an atmosphere containing 5-10% CO2 at 37 degree C and identified through biochemical and fermentation tests. A Lasota like virus was also isolated from the same material which showed HA activity but was found negative to Avian Influenza virus against known Avian Influenza serum. The isolated virus was sent to Central Veterinary Laboratory, Weybridge, U.K. which confirmed it to be Lasota virus having comparatively high Intracerebral Pathogenicity Index (ICPI) as 0.90. Attempts for the transmission of the disease in susceptible healthy broiler chicks was successful through aerosol route using combination of inoculum of the isolated bacteria and virus. On post-mortem of infected birds air- sacculitis, tracheitis and unilateral pneumonia were noticed. Again ORT and NDV were isolated from the material harvested from these chicks. According to the observations of these experiments, it was concluded that the isolated ORT had a triggering effect on Lasota virus or vice versa. The environmental temperature and other stress factors might have aggravated the disease problem.

This study was conducted to examine comparative productive performance of different varieties of Desi chickens identified from group of Desi flock procured from rural field conditions of Mianwali and Bhakkar districts of Punjab province (Pakistan). A total of five varieties of Desi chickens viz., dark brown, medium brown, light brown, black and mixed colour were identified on the basis of their plumage colour and uniformity of body characters. Four hundred and eighty birds (96 of each variety) were selected for further study. The birds of each variety were randomly divided into 3 equal replicates. They were maintained in 15 separate pens on deep litter system under optimal managemental conditions and were fed a balanced layers ration ad-libitum. The birds were provided 16 hours light per day. The experiment was conducted for 8 weeks. The results indicated that varieties differed significantly in egg production, feed conversion efficiency and egg mass, whereas, no difference was observed in feed intake and egg weight between them. Based on the findings of this study, it appeared that different varieties of Desi chickens identified and selected during the course of this study on phenotypic basis differed in their productive potential on the basis of better productive potentials in terms of egg yield, FCR and egg mass, dark brown, and black varieties of Desi chickens could be used to carry out further selective breeding work to study possibility of improving their productive potentials.

Objective: The study was conducted to investigate the gut content and record morphometric and hematological parameters in the common hoopoe (Upupa epops). Materials and Methods: Twenty samples of healthy birds (10 from each sex) were collected from different locations in Okara District, Punjab, Pakistan, from September 2020 to March 2021. Birds were captured live for blood samples and morphometric and gut analyses. Results: It was revealed that the concentrations of different hematological parameters were as follows: hemoglobin, 20.03g/dl; red blood cells, 3.28 × 106/μl; white blood cells, 326.67 × 103/ μl; hematocrit, 56.47%; MCV, 173.33 FL; MCH, 57.4 pg; MCHC, 57.4 pg; PLT, 8.33/μl; and RDW, 8.33/μl. The percentages of neutrophils, lymphocytes, monocytes, and eosinophils were 84.67%, 11.67%, 2.00%, and 1.67%, respectively. The gut content of the common hoopoe mostly consisted of Coleoptera and Acrididae larvae. However, Lepidoptera, Gryllotalpidae, and sand were also recorded, along with seeds of Salvadora persica. Conclusions: There were no significant differences between male and female U. epops in feeding content, total weight of the gut, or weight of the empty gut. Regarding the morphometric param¬eters, there was a significant difference in both sexes' wingspan, body length, and body weight. Males were significantly heavier than females. [J Adv Vet Anim Res 2022; 9(2.000): 290-294]