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Present study was designed to prepare and evaluate gel based combined vaccine against Peste des Petits Ruminants (PPR), Contagious Caprine Pleuropneumonia (CCPP), Foot and Mouth Disease “O serotype” (FMDO) to mitigate the cost and frequency of the vaccination. Gel based vaccines contain either single immunogen (monovalent) of PPR, CCPP or FMDO, two (bivalent) PPR+FMDO, PPR+CCPP, FMDO+CCPPP or three (trivalent) PPR+CCPP+FMDO. Each dose (0.5 ml) of either of the vaccines contained 105 tissue culture infective dose (TCID50) units of FMDO virus, 107 TCID50 units of PPR virus and 0.15 mg of Mycoplasma mycoides capri. Each of the vaccine was injected subcutaneously to each of the goats (n=4). Antibody response of goats to FMDO, PPR and CCPP was determined by complement fixation test (CFT), virus neutralization test (VNT) and enzyme linked immunosorbant assay (ELISA), respectively at 0, 60, 120 and 180 days post-priming. Antibody response of the goats to either of the immunogen was not significantly different irrespective to the form of vaccine (monovalent, bivalent or multivalent vaccine) (P>0.05). It indicated that either of the immunogen did not interfere the immunogenesis process of other immunogens in the same vaccine. [ABSTRACT FROM AUTHOR]

Auxin, one of the most important phytohormones enhances the root architecture, nutrient acquisition and plant growth. Present study provides the screening, characterization and optimization of auxin producing bacteria from wheat rhizosphere. Effects of auxin producing isolates on seed germination and growth of T. aestivum were also evaluated. The Bacillus megaterium, Escherichia coli, Klebsiella pneumonia and Bacillus marinus (AUX36, AUX53, AUX137 and AUX142 respectively) were selected by salkowski reagent as auxin producers. Colorimetric analysis showed that all selected isolates produced maximum auxin in the presence of tryptophan at 37 °C and pH 7 after 72 hours of incubation. AUX36 and AUX137 produced maximum auxin at 0.98% (w/v) NaCl concentration, while AUX53 and AUX142 at 2% NaCl. Glucose supplementation enhanced the auxin in the culture filtrate of AUX53 and AUX137, while addition of sucrose in the growth medium enhanced the auxin production for all isolates except AUX142. The AUX36, AUX53 and AUX137, produced highest concentration of auxin at 0.1% peptone. Seed germination experiments using surface sterilized seeds of T. aestivum revealed that all isolates could increase root length of seedlings as compared to control. Comparison of various growth parameters after 15 days of growth showed that plant growth was significantly improved by bacterial treatments. All isolates had significant effect on root and shoot length of T. aestivum. It is concluded that Bacillus megaterium AUX36, Escherichia coli AUX53, Klebsiella pneumonia AUX137 and Bacillus marinus AUX142 may be used to develop indigenous biofertilizers in future after further investigations. [ABSTRACT FROM AUTHOR]

Summary: 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. [ABSTRACT FROM AUTHOR]

Newcastle disease virus (NDV) live vaccines are supplied in lyophilized form and usually administered through conventional routes (drinking water, spray, or eye drop) following reconstitution in a diluent. Virus inactivation due to physico-chemical properties of the diluent at the time of administration may lead to vaccine failure. The present study aimed to evaluate the survival of NDV live vaccine strain immersed in 5 pH-amended water samples (pH 5.00, pH 6.00, pH 7.00, pH 8.00, and pH 9.00) by sequential determination of virus infectivity on Vero cells for 3 hours. Minimum reduction in virus infectivity was recorded in the water with neutral or slightly alkaline pH, while the virus was relatively less stable at extreme pH conditions. Maximum reduction of infectivity was observed in the water with pH 9.00 in which the virus was completely inactivated within 3 hours. Addition of stabilizers (Cevamune® or skimmed milk) slightly altered the pH and total dissolved solids (TDS) values of the virus-charged water samples. In the stabilizer-added water samples, minimum reduction in infectivity was observed in the water with neutral pH, followed by the ones with a pH of 8.00, 6.00, 5.00, and 9.00. In all types of water samples, T-90 values (time required for 90% reduction in virus infectivity) were highest (485 minutes) at neutral pH (pH 7.00) and lowest (102 to 134 min) at an extreme alkaline condition (pH 9.00). Results of the present study indicate that water with a pH range of 7.00 to 8.00 is suitable for administration of NDV live vaccines. However, the addition of Cevamune® or skimmed milk may have beneficial effects on preserving the infectivity of the virus, even at extreme pH conditions. [ABSTRACT FROM AUTHOR]

Colchicine is microtubule inhibiting agent used for thecure of various skin ailments causing inflammation and reduction of pain in gouty arthritis.The current study was designed to evaluate the effect of colchicine on cellular immune responses in mice. Cellular immune responses were assessed indelayed type hypersensitivity (DTH) assay, macrophage engulfment assay, cyclophosphamide induced neutropenic test and nitric oxide (NO) production assay.Colchicine was administered to mice at doses of 40μg/kg, 80μg/kg and 160μg/kg intraperitoneally. Data were statistically analyzed by ANOVA and chi square test. DTH showed significant (P<0.0001) decrease in skin thickness of colchicine treated groups. In macrophage engulfment assay, significant (P<0.0001) decrease in engulfment of sheep red blood cells (SRBCs) was observed by macrophages in colchicine treated groups. Colchicine treated groups showed significant (P<0.0001) reduction of TLC and neutrophil count.NO production difference was significantly (P<0.0001) reduced in macrophages alone and lipopolysaccharide (LPS) stimulated macrophages in colchicine treated groups. Therefore, it is concluded that colchicine suppresses the cellular immune responses in mice. [ABSTRACT FROM AUTHOR]

This study was planned to investigate effect of colchicine on humoral immune responses and immune organs in mice. Humoral immune responses were evaluated by performing haemagglutination assay, mice lethality test and Jerne hemolytic plaque formation. Colchicine treated groups were administered 40μg/kg, 80μg/kg and 160μg/kg.Data obtained was analyzed by ANOVA. There was significant(P <0.0001)reduction in HA titer of colchicine treatedgroups. With the increasing dose, there was reduction in the HA titer. Colchicine enhanced the mortality after administration of bovine Pasteurellamultocidaculture. In addition, there was significant (P<0.0001) reduction in body weight and spleen weight difference of mice in colchicine treated groups. There were no histopathological changes observed in spleen and thymus at 40μg/kg and 80μg/kg doses of colchicine. At 160μg/kg dose, increase in thickness of trabecular was seen due to edema in the spleen. This study has shown that humoral immunity is suppressed by the administration of colchicine. [ABSTRACT FROM AUTHOR]

Neonatal calf diarrhea is one of the main economic losses to dairy industry in developing countries. Study was planned to assess the prevalence of rotavirus diarrhea in Pakistan. Fecal samples (n=200) from cattle and buffalo diarrheic calves, age<3months were collected aseptically from ten districts of Punjab province selected on the basis of agro-ecological zones (canal fed, rain fed, desert and Pothohar).Commercially available ELISA kit was used for rotavirus detection. Results revealed 6% positivity including 3% from cattle calves and 4% from buffalo calves. District wise analysis showed higher prevalence(4%) in Lahore as compared to Faisalabad district(2%). Statistically significant (P<0.05) correlation was found between diarrhea and age groups of calf showing<1month susceptible for diarrhea, similarly significant association was found among gender, housing type, feeding pattern and season. We could not establish any correlation between species. Statistical analysis showed significant correlation between diarrhea incidences and source of water as canal water, underground and tap water. The present findings revealed rotavirus infection in calves in Pakistan that needs to be addressed using vaccine and improving livestock management. [ABSTRACT FROM AUTHOR]

Phytase is an enzyme which breakdown the complex organic (unavailable) forms of phosphorous such as phytic acid into simpler inorganic (available) forms. Aim of the present study was to isolate, characterize and optimize physicochemical conditions of phytase producing bacteria. Phytase producing bacterial isolates were screened on phosphate solubilizing media. Isolates (PHY02, PHY03, PHY06, PHY07, PHY12, and PHY30) were identified by morphological, microscopic, and biochemical characteristics as Lactobacillus casei, Enterobacter intermedius, Bacillus badius, Escherichia coli, Shigella spp., and Klebsiella spp., respectively. All isolates (PHY02, PHY03, PHY06, PHY07, PHY12, and PHY30) produced optimum level of phytase on 37°C (27, 9, 19, 40, 19, and 32 IU/ml, respectively), pH 6.5 (26, 15, 19, 41, 19, and 32 IU/ml, respectively) and 1% NaCl (27, 15, 17, 41, 18, and 32 IU/ml, respectively). PHY03, PHY06, PHY07 and PHY12 had significantly higher (P<0.05) enzyme production when glucose was used as sole source of corbon, while 0.3% lactose was preferred by PHY02 and PHY30. Peptone was preferred nitrogen sources for PHY02, PHY12, PHY07 and PHY30, while tryptone was preferred by PHY03 and PHY06, respectively. Phytase producing bacterial isolates may be further analyzed and developed as indigenous biofertilizers. [ABSTRACT FROM AUTHOR]

Present study has been designed to check the pathological effects of a low pathogenicity H9 avian influenza virus on broilers following experimental infection through intranasal, ocular and oral route at 21 days of age. Moderate clinical signs were observed which started appearing at two days post infection (PI), became severe on five days while started subsiding from seven days onwards. Severity of the signs and lesions was highest in respiratory organs following intranasal route of infection. No mortality was observed except one bird each in the groups infected through intranasal and oral route of infection. Histopathology of the organs in the birds infected through intranasal route revealed severe hyperemic trachea and congested lungs. While moderate congestion of kidneys and mild necrosis in the liver tissue was seen in all treatment groups. Detection of virus antigen through immunohistochemistry was possible in trachea, lung, and kidney of few birds on two days PI while in all birds at five days PI. In some of the birds inoculated through either of the route, the virus was detected in liver and duodenum tissue at five days PI. [ABSTRACT FROM AUTHOR]

The article discusses the results of a comparative diagnostic study on the applications of antigen-capture enzyme-linked immunosorbent assays (AC-ELISA) and immunohistochemistry (ICH) to detect bovine viral diarrhea (BVD). The research uses a serum and ear notch biopsy among bovine to determine its persistent infection. It conclude that AC-ELISA and ICH were equally suitable for detection of BVDV persistent infection.

Small ovarian size and low number of follicles are major causes of low reproductive potential in buffalo (Bubalus bubalis). There is no tool through which reproductive performance of buffalo can be phenotyped. Therefore, aim of the present study was to develop a method on the basis of which reproductive performance of buffaloes can be phenotypically assessed. For this purpose two experiments were designed. In first experiment, ten Nili-Ravi buffalo heifers were used in the study to measure repeatability of follicular count. Each heifer was scanned on alternate day, from day 1 to day 9 of the successive estrous cycle. Antral follicles and repeatability was assessed in different follicular waves. In 2nd experiment, 4 lactating buffaloes were selected to count antral follicles from day 1 to day 10 of the estrous cycle and super-stimulated by using FSH hormone. Follicular count after FSH injection was examined by ultrasound. Follicular count is highly repeatable in different follicular waves within (r = 0.83) and between (r = 0.85) different estrous cycles. Follicular count and super stimulatory response are positively correlated (r = 0.95). In conclusion, animals with high antral follicular count can be selected for super stimulation and embryo transfer to get optimum response. [ABSTRACT FROM AUTHOR]

Pesti des Petits Ruminants (PPR) is an acute contagious viral disease of small ruminants and is causing heavy economic losses to goat industry. It is controlled by mass vaccination and bio-security measures. In present study, in process quality control factors such as variable biological titer, different adjuvants and shelf life of the PPR vaccines affecting antibody response of the goats were evaluated. The freeze dried vaccine with a biological titer of 10[sup 5.00] TCID[sub 50] per dose provoked maximum antibody titer followed by the ones with a titer of 10[sup 4.00] or 10[sup 3.00]TCID[sub 50] which provoked nearly equivalent antibody response while the goats inoculated with a vaccine having 10[sup 2.00] TCID[sub 50] virus concentrations developed minimum antibody titer. The oil based PPR virus vaccines elicited significantly higher antibody response (p<0.05) while gel based vaccines induced relatively less antibody titer but freeze dried PPR vaccine induced minimum antibody titers. Each of the adjuvant vaccines (oil or gel) and non- adjuvant vaccine containing biological titer of 10[sup 4.00] TCID[sub 50] induced more than 50 Percent Inhibition (PI) values (protective antibody titer). The freeze dried vaccine stored at 4 °C did not show any significant drop in the biological activity of the virus even after 12 months of the storage. Moreover, both the freeze dried and adjuvant vaccines when stored for 12 months at 4 °C did not show any effect on their antibody response in the vaccinated goats. It is concluded that oil adjuvant vaccine with 10[sup 4.00] TCID[sub 50] units of biological titer of virus induced the highest antibody titer in goats that persisted for more than 10 months post priming. [ABSTRACT FROM AUTHOR]

The interaction of nickel nanoparticles with Escherichia coli (E. coli) bacteria has been studied. The nickel nanoparticles have been fabricated by continuous wave laser ablation of nickel target and their properties are studied using different characterization techniques. The antibacterial activity of nickel nanoparticles was checked against E. coli bacteria. Escherichia coli were cultured in nutrients broth and different concentrations of nickel nanoparticles were added to bacterial culture solution to investigate the interaction of nickel nanoparticles with bacteria and to check toxicity of the nickel nanoparticles against E. coli. The fabricated Ni nanoparticles have exhibited considerable antimicrobial activity against E. coli. [ABSTRACT FROM AUTHOR]