RECENT UPDATES ON MOLECULAR DETECTION OF H9N2 AS LOW PATHOGENIC STRAIN OF AVIAN INFLUENZA VIRUS FROM POULTRY FARMS OF LAHORE, PAKISTAN

H9N2 avian influenza outbreaks have caused great economic losses to the poultry industry in recent decades due to a decrease of egg production, high morbidity, and mortality. Due to different antigenic variants, Influenza virus has become problematical because it has the ability to cross the species barrier. As it is highly pathogenic so its diagnosis and vaccines are of high importance. Hemagglutination inhibition (HI) test is mostly used for subtyping and detection of antibody titer against the virus. Furthermore, its continuous mutations in the HA gene transforms AIV subtype H9N2 (a low pathogenic subtype) into high pathogenic virus subtypes like H5N2 and H7N7 that may have pandemic potential. Thus, it is necessary to identify various antigenic variants of Influenza virus, so it is direly needed to study the HA gene, its attachment to host receptors, the release of genetic material and pathogenicity. In the present study, virus samples from poultry were isolated. Both serological and molecular confirmation was done for 100 samples collected from the different area. They were properly labeled and prepared for the process of egg inoculation in embryonated eggs. The virus was grown in amnioallantoic membrane of embryonated eggs and harvested fluid is then proceeded for confirmatory testing. Hemagglutination and Hemagglutination inhibition testing was done. RNA was extracted by the kit method and cDNA was synthesized. Reverse transcriptase (RTPCR) was performed using specific primer sets and then the PCR product was run on agarose gel. The bands obtained were sent for sequencing.

CANINE TESTICULAR HISTOPATHOLOGY ENSUING CHEMICAL CASTRATION

Castration in domestic and companion animals plays an important role in controlling animal population, aggressive male behaviour and producing meat with a consistent quality acceptable to consumers. Surgical castration is the widely used method, but it is more expensive and painful if not well managed. This research focused on the use of 10% iodine and 1% polidocanol foam as intratesticular injections to achieve castration in three adult dogs owned by a client in Lwandeti village in Kakamega County, Kenya. Two of the dogs received 1ml of 1% polidocanol, while one dog received 1ml of 10% iodine in each of their testicles. The testicles were surgically harvested after three months for histological examination. The histological examination revealed that the seminiferous tubules of testes those received 10% Lugol’s iodine had cellular infiltration and fibrin deposition. Those that received 1% polidocanol showed moderate to severe cellular infiltration but less than the iodine group. This study has shown the potential of polidocanol and iodine as chemical methods that can be used instead of surgical castration.

PHYTOCHEMICALS ANALYSIS AND ANTIMICROBIAL ACTIVITIES OF ECHINOPS ECHINATUS FROM CHOLISTAN DESERT, PAKISTAN

Plants have been used as a source of food as well as medicines by the man throughout history. Echinops echinatus was evaluated for phytochemicals and antibacterial activities against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia and Anti-urease inhibition assay activities were tested during the study. The laminar flow cabin was cleaned with 70% ethanol. α-glucosidase inhibition activity was performed according to the standard method with slightly modification. Carbohydrates, phenols and tannins were found positive in leaves, stems and roots. Saponins and glycosides were found in flowers, leaves, roots and stems. While roots and stems revealed the presence of steroids. Terpenoids were found in flowers and leaves. The leaves showed the maximum inhibition zone 10mm against K. pneumonia while against E. coli, S. aureus and P. aeruginosa was found inactive. K. pneumoniae, E. coli and P. aeruginosa were resistant to flowers extract. The root extract showed maximum zone of inhibition 18 mm against S. aureus while against P. aeruginosa and K. pneumonia inhibition zone was 15mm and 14mm respectively. The extracts of leaves, flowers, roots and stem showed positive results for α-glucosidase. The roots extract showed maximum inhibition with 75.3±1.5 with IC50 value of 207.3±1.3 for α-glucosidase. Urease inhibitory activity of stem extracts showed maximum inhibition activity 93.71±0.86 with IC50 value 15.63±0.42. While leaves and roots showed almost same activity 92.45±0.63 and 92.63±0.76, respectively. Echinops echinatus could be considered for further studies in the treatment of various ailments as a natural remedy.

BIOLOGY AND ECOLOGICAL ADAPTATIONS OF BLACKBUCK (Antilope cervicapra): A REVIEW

Blackbuck is the existing members of genus Antilope which is most elegant and graceful among all Antelopes of Asia with distinct sexual dimorphism. Blackbuck show endemism in Pakistan, Nepal and India occupying mainly the semi-arid grassland areas. The name of species attributes towards the dark brownish to blackish coat color of male species. While female and the young ones are tawny or yellow. Chin undersides of legs and chest are white in both male and female individuals. Males have whorled horns with 79cm length with absence in females. The average body length of this animal is 100-150cm with the tail length of 10-17cm. The average body weight for male is 20-57kg and for female 19-33kg. Thin grassy forests, open and semi-desert areas are good habitat for it. Being diurnal and herbivorous, it acts as both grazers and browsers. It is the fastest animal with an average speed of 80km/h. It remains reproductively functional whole year. Reproductive disorders including dystocia and different infectious diseases due to ectoparasites, endoparasites, bacteria and viruses affect these species. Habitat loss, stress, illegal killing and genetic troubles cause the species to be ‘Extinct in the Wildlife’ in Pakistan so conservation strategies are underway for species protection. It is indispensable for zoologists and conservational biologists to observe the species for its conservation and confronting threats. Review article highlighted necessary information about species, which will clear the way for further research on species.

EFFECT OF TOCOPHEROL AND SELENIUM ON THE PERFORMANCE OF BUSERELIN FOR ESTRUS INDUCTION IN LATE SEASON ANESTRUS MARES (EQUUS CABALLUS)

The aim of this study was to determine the effect of tocopherol and selenium on the performance of buserelin (GnRH analogue) for estrus induction in anestrus mares. A total of 18 seasonally anestrus mares were divided into three equal groups (n=6 in each) after ovarian scanning. Mares in group A (treatment group) were treated with tocopherol and selenium intramuscularly along with daily dose of buserelin. Mares which came into heat within ten-day buserelin protocol, further injections of buserelin for those mares were stopped. Mares in group B were treated with buserelin intramuscularly only for ten days. Mares which came into heat within ten-day buserelin protocol, further injections of buserelin were stopped. Blood/serum was collected from each experimental mare kept in different groups (A-C) at days 1, 11, 12, 13 and 14. Onset of estrus signs was 83.3, 33.3 and 0% in group A, B and C, respectively, that was significant (P<0.05) among groups. Follicular dynamics were 20.24±3.00, 14.59±2.24 and 10.35±0.63mm in groups A, B and C, respectively with significant (P˂0.05) difference. Change in serum E2 contents was significantly (P˂0.05) different among groups A, B and C whereas progesterone concentration showed non-significant difference among groups.

PHENOTYPIC, GENETIC AND ENVIRONMENTAL CORRELATIONS BETWEEN BODY WEIGHT AND LINEAR BODY TRAITS OF CHICKEN GENOTYPES

Data from 123, 49, 116, 137, 42 and 64 chickens belonging to Isa Brown x frizzle feathered (IBxF), Isa Brown x naked neck (IBxNa), Isa Brown x normal feathered (IBxN), frizzle feathered x Isa Brown (FxIB), naked neck x Isa Brown (NaxIB) and normal feathered x Isa Brown (NxIB) genotypes, respectively were used to establish phenotypic (rP), genetic (rG) and environmental (rE) correlations between body weight and linear body traits at 2, 10 and 20 weeks of age. Phenotypic correlations were analyzed by Pearson’s Product Moment method. Genetic and environmental correlations were estimated by covariance analysis. The highest rP estimates were obtained between body weight and shank length (SL) in IBxF (0.84) and NxIB (0.87) at 10 weeks, body width (BW) in IBxNa (0.84) and IBxN (0.72), drumstick length (DL) in FxIB (0.71) and NaxIB (0.88) at 20 weeks. Body weight had highest rG estimates with SL in NxIB (0.99), BW in IBxF (2.40), wing length (WL) in BxNa (0.91) and FxIB (7.39) at 10 weeks, body girth (BG) in IBxN (0.97) and BW of NaxIB (4.73) at 20 weeks. However, SL of IBxNa (-5.15), BG of NaxIB (-0.53), keel length (KL) of NxIB (0.35), WL of IBxF (-0.38) and FxIB (-1.41) at 2 weeks and WL of IBxN (-0.43) at 20 weeks had the least rE with body weight. Indirect selection of body weight using the highest phenotypic, genetic and lowest environmental correlated linear traits will result in rapid genetic improvement in these chickens.

ISOLATION, CHARACTERIZATION AND GENOME ANALYSIS OF A NOVEL VIRULENT ESCHERICHIA COLI BACTERIOPHAGE VB_ECOM_011D4

Phage therapy is a promising alternative therapy for the treatment of E. coli infection. Although the total number of phages on the earth is as high as 10 31 , the reported phages and thoroughly studied are very limited. Therefore, the continuous discovery of new phages and in-depth research will provide materials for the wide application of phage therapy in the future. In this study, a novel E. coli phage vB_EcoM_011D4 was isolated from sewage samples, and the biological characteristics were studied. Electron microscopy and homology analysis results showed that vB_EcoM_011D4 belongs to the family Myoviridae. One-step growth curve showing the latent period of vB_EcoM_011D4 was 10 min, with the burst size of 115 PFU/cell. Additionally, Phage vB_EcoM_011D4 was highly stabled under different temperatures (range 4 – 70 ℃) and pH conditions (range 6 – 10). At the same time, its genome was subjected to high-throughput sequencing and compared with the reported phages. The results of high-throughput sequencing assembly showed that vB_EcoM_011D4 is a linear, double-stranded DNA virus containing 163764 bp, with an average GC content of 40.50%, and a total of 273 open reading frames (ORFs). Genomic comparison analysis revealed that most of the ORFs were similar to Enterobacteria phage Phi1 and RB49. However, ORF147 and ORF148 putative DNA methylase family protein is less than 67% homology with already published phages. In addition, the phylogenetic analysis of terminates large subunit showed that it belongs to a new branch and shows less than 50 similarities to reported phages. There is no lysogenic, toxin or antibiotic-resistant related gene was found in the genome of vB_EcoM_011D4. In summary, vB_EcoM_011D4 is a newly discovered phage, which can be further studied for elucidating the phage diversity and it is benefits for the wide application of phage therapy.

BIOFORTIFICATION OF HORTICULTURAL CROPS WITH SELENIUM

Selenium (Se) is a naturally occurring element, structurally resembling with Sulphur, lying in group 6 of the periodic table. It is an essential micronutrient, required for proper functioning of many biological processes like hormone formation, immune system, hair growth, muscular movements and reproduction and helps in defense against many diseases like viral infections, Keshan’s diseases and arthritis, as it is important component of selenoproteins and enzymes like glutathione peroxidase. Low selenium level in human body can lead to various diseases and disorders. To fulfill selenium dietary intake, selenium enriched diet is necessary. Plants are the main source of human diet. Plants are fortified with selenium by adding different forms of selenium through different methods. They uptake or absorb selenium through Sulphur assimilation pathway and store it into the tissues in either organic or inorganic form. Till now much research has been carried out in production of se-fortified crops. This review paper is aimed at reviewing recent studies which have been done on biofortification of horticultural plants with selenium, like fruits and vegetables, which contribute a major portion of human diet.

DIAGNOSIS AND TREATMENT OF A CAT WITH IMMUNE MEDIATED HEMOLYTIC ANEMIA (IMHA)

In order to explore the diagnosis and treatment of immune-mediated hemolytic anemia (IMHA) in cats, we diagnosed and treated a cat with IMHA. We made a detailed observation and record of the treatment process for reference to the treatment of IMHA in cats. Our diagnosis was based on the cat’s clinical manifestations, routine diagnosis, laboratory tests, and ultrasound results. We excluded the possibility of common infectious diseases and parasitic infections. The cat was initially diagnosed with hemolytic anemia, fatty liver, hepatocyte injury and cholestasis. The condition did not improve after symptomatic treatment for five days. But the cat gradually improved after the use of immunosuppressants, and was finally diagnosed to be suffering from IMHA. After 32 days of treatment, the cat was basically cured and discharged from the hospital. This paper describes a case study which can serve as the reference for the diagnosis and treatment of feline IMHA.

HYPOGLYCEMIC EFFECT OF FLAXSEED LIGNAN AND β GLUCAN IN POSTMENOPAUSAL DIABETIC PATIENTS

Diabetes mellitus is an important metabolic disorder and owing to its significant health impacts is a leading cause of death around the globe. According to WHO estimates, Pakistan is the 7th highest diabetes affected population of the world where almost 7 million peoples are suffering from the disease. Flaxseed lignan and beta glucan have constructive role in controlling diabetes. In connection to their enhanced insulin sensitivity, dietary supplementation of flaxseed fiber has shown potential results in postmenopausal diabetic women. The subject population was feed with lignan and beta glucan capsules and hypoglycemic activity was monitored both on daily and weekly basis. Blood samples was taken to observe the blood glucose by use of flaxseed lignan and β glucan in postmenopausal women. Fasting and random glucose levels of postmenopausal diabetic women in 4 weeks fed on flaxseed lignan and β glucan (P<0.01). Liver and kidney functioning tests are found to be non-significant. This research indicates that flaxseed lignan and β glucan reduces the random and fasting blood glucose levels in postmenopausal patients.