Effect of induced Fasciola gigantica infection during pre-patency on the performance of buffalo calves fed on different percentage of protein.

Thirty growing Murrah buffalo calves (8-12 months of age, 109.85 ± 2.43) were reared in parasite free conditions and randomly divided into three equal groups as per CRD. They were fed on iso-caloric (2.01 ME Mcal/Kg diet) diets containing standard protein (SP) diet at 100 %, 90 % of SP (medium protein, MP) and 80 % of SP (low protein, LP) of the protein requirements (Kearl 1982). After 21 days of feeding, each group was further subdivided into two sub-groups (A & B). Animals in sub-groups 'A' served as non-infected control, while in sub group 'B' were orally infected with Fasciola gigantica metacercarie (mc; 1,000 each). A metabolic trial of 40 days post infection was carried out in control and parasitized animals. Intake of digestible dry matter, organic matter and acid detergent fibre (ADF) was significantly higher (P < 0.05) in SP group compared to LP group. The digestibility of crude protein (CP) and ADF was significantly higher in SP group compared to MP and LP groups. The digestible crude protein (DCP) and total digestible nutrients (TDN) intakes (g/kgW(0.75)) were also significantly (P < 0.001) higher in SP than MP and LP groups. However, DCP intake was significantly (P < 0.001) lower in infected subgroups compared to control subgroups. Intake and balance (g/d) of nitrogen, calcium and phosphorus were significantly (P < 0.01) higher in SP than MP and LP groups. The average daily gain of buffalo calves fed on SP Uninfected (SPU), SP Infected (SPI), MPU, MPI and LPU, LPI groups was 333, 178, 356, 144, 222 and 144 g and was significantly (P < 0.01) lower in animals fed LP ration. The feed conversion ratio (FCR) was also significantly (P < 0.01) higher in infected sub-groups as compared to respective control groups. The results showed that a SP diet substantially improved the overall performance of buffalo calves in control and infected groups and reduced adverse effect of F. gigantica infection.

Interferon-gamma and interleukin-4 expression during Fasciola gigantica primary infection in crossbred bovine calves as determined by real -time PCR.

Interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) expression in crossbred (Bos taurusxBos indicus) bovine calves during primary infection with Fasciola gigantica was measured. Ten crossbred calves of 1-year age were divided into two groups of five calves each, group I uninfected control and group II calves orally infected with a dose of 1000 metacercariae of F. gigantica. The two cytokines were measured 10, 30 and 75 days post-infection (PI) by real-time polymerase chain reaction (RT-PCR) with the double stranded DNA-binding dye SYBR Green. IL-4 was present in detectable levels in peripheral blood mononuclear cells (PBMCs) of infected animals at 10, 30 and 75 days PI but no IFN-gamma was detected in PBMCs of infected animals at 10 and 30 days PI. However, at 75 days PI, IFN-gamma in two infected animals was present in detectable level. Eosinophil count increased from 2nd fortnight after infection and the increased level persisted till the termination of experiment. Present study indicated that T-cell response during F. gigantica infection was Th2-type during earlier phase of infection, which may be polarized in chronic infection to that of a Th0-type.

Improved HPLC method for the simultaneous determination of allantoin, uric acid and creatinine in cattle urine.

An HPLC procedure developed for the rapid and simultaneous determination of purine derivatives (PD) in ruminants' urine was investigated, since the adoption of a single method for the simultaneous detection of PD and creatinine was not carried out due to elution of polar co-extractives and also due to overlapping of the peaks of allantoin and creatinine. The experimental conditions chosen in the present study avoid the presence of chemically competitive compounds and afford a good separation of the peaks of allantoin and creatinine. The recoveries of the standard compounds added to urine samples were 94-104%. This method can be proposed as a possible reference method for the estimation of allantoin, uric acid and creatinine in cattle urine.

Novel scintillation proximity assay for measuring membrane-associated steps of peptidoglycan biosynthesis in Escherichia coli.

We have developed a novel, high-throughput scintillation proximity assay to measure the membrane-associated steps (stages 2 and 3) of peptidoglycan synthesis in Escherichia coli. At least five enzymes are involved in these two stages, all of which are thought to be essential for the survival of the cell. The individual enzymes are difficult to assay since the substrates are lipidic and difficult to isolate in large quantities and analysis is done by paper chromatography. We have assayed all five enzymes in a single mixture by monitoring synthesis of cross-linked peptidoglycan, which is the final product of the pathway. E. coli membranes are incubated with the two sugar precursors, UDP-N-acetyl muramylpentapeptide and UDP-[(3)H]-N-acetylglucosamine. The radiolabel is incorporated into peptidoglycan, which is captured using wheat germ agglutinin-coated scintillation proximity assay beads. The assay monitors the activity of the translocase (MraY), the transferase (MurG), the lipid pyrophosphorylase, and the transglycosylase and transpeptidase activities of the penicillin-binding proteins. Vancomyin, tunicamycin, nisin, moenomycin, bacitracin, and penicillin inhibit the assay, and these inhibitors have been used to validate the assay. The search for new antimicrobial agents that act via the late stages of peptidoglycan biosynthesis can now be performed in high throughput in a microtiter plate.

Primary experimental infection of riverine buffaloes with Fasciola gigantica.

The clinical course of the primary experimental Fasciola gigantica infection was investigated in riverine buffalo calves of the Murrah breed. Nine male calves aged 12-15 months were randomly assigned to two groups of five (Group I) and four (Group II) animals. Each animal in Group I, was orally infected with 1000 metacercariae (mc) of F. gigantica, whereas Group II animals did not receive any infection dose and served as uninfected controls. No clinical signs of fasciolosis were observed until the sixth week post-infection (PI). Group I animals, however, developed recognised symptoms of acute fasciolosis, comprising apyrexic inappetance, anemia, poor weight gain, diarrhoea and sub-mandibular and facial oedema, respectively, from 5, 6, 8, 16 and 17 weeks PI. The signs were intermittent in nature and of variable duration. The prepatent period was of 92-97 days (mean 95.2 +/- 3.1). One of the five infected animals died on Day 147 PI. At necropsy, 36.8 +/- 11.0% of the infection dose was recovered as adult fluke population. The gross lesions were primarily biliary in nature. Group II, the uninfected controls, throughout the study period of 165 days PI, did not show any symptom and were negative for F. gigantica. The study demonstrated that the onset of adverse effects of F. gigantica on the growth and health of the infected host was mainly noted during late prepatency much before coprological prediction and diagnosis. The significance of preventive therapy against fasciolosis during prepatency has been stressed in endemic areas.

Effects of Fasciola gigantica infection on growth and nutrient utilisation of buffalo calves.

The effects of Fasciola gigantica infection on bodyweight gain, dry matter intake, digestibility of nutrients and feed conversion efficiency in buffalo calves were investigated. Nine male buffalo calves of the Murrah breed, aged 12 to 15 months with a mean (se) bodyweight of 166 (12.5) kg, were randomly assigned to groups of five (group 1) and four (group 2). The animals in group 1 were given 1000 viable, mature metacercariae of F gigantica orally, while the animals in group 2 served as uninfected controls. They were stall fed on diets containing a concentrate mixture and ad libitum wheat straw and were maintained by standard management practices for a period of 165 days after infection. The average daily liveweight gain of the infected animals was 110.6 g, compared with 439.4 g in the uninfected controls, and was associated with the appearance and establishment of immature flukes in hepatic bile ducts. The feed conversion efficiency declined significantly (P<0.01) from 41 days after infection and was lowest at the end of the experiment. F gigantica infection did not influence the digestibility of the nutrients. The impaired feed conversion efficiency was mainly due to a reduction in dry matter intake due to inappetence.

Possibility of using 131I-albumin as a marker for the estimations of microbial protein synthesis rates in the rumen.

Total microbial protein synthesis rates in the rumen of buffaloes were estimated by isotope dilution technique, using 131I-albumin treated with tannic acid as a marker. The animals were fed groundnut cake treated with formaldehyde to meet 50% of their digestible crude protein (DCP) requirement and 2.5% urea molasses mixture was given to meet the remaining requirement of DCP. Wheat straw was fed as the basal roughage. The total average microbial protein synthesis was 58.14 g/day.

Comparison of the production rates of bacteria in the rumen estimated by using labelled live and formaldehyde treated mixed bacterial cells.

The bacteria production rates in the rumen have been estimated by injecting 14C- and 35S-labelled mixed rumen bacteria, either live or killed by treatment with formaldehyde, into the rumen and applying isotope dilution technique. The rate of bacteria production when estimated by using either live- or dead-(protected-)labelled bacterial cells were comparable.

Comparison of colposcopy directed biopsies and cold knife conization in patients with abnormal cytology.

One hundred patients with abnormal cytology were evaluated by colposcopy, and histologic material was obtained by directed biopsies, cold knife conization and endocervical curettage. The colposcopy directed biopsies provided the diagnosis in 94 per cent of the patients, and the colposcopic impression was accurate in 89 per cent of the patients. Cold knife conization was performed in 98 patients, since two patients had invasive carcinoma on colposcopic directed biopsy. In 75 patients, the histopathologic finding on cone biopsy was in complete agreement with the directed biopsies. In 11 patients, the directed biopsies were a degree higher than on cone biopsy, and in six patients the histopathologic findings were a degree lower. In six patients the colposcopic directed biopsies missed the diagnosis of dysplasia. No patients with carcinoma in situ or invasive carcinoma were missed by the colposcopic directed biopsies.

Pregnancy with bilateral ruptured benign cystic teratomas.

A case of intra-abdominal rupture of bilateral cystic teratomas in the first trimester of pregnancy is reported. Review of the literature revealed 22 cases of intraperitoneal rupture of benign cystic teratoma (1908 to 1971). In all of these cases only one ovary was involved, and the period of gestation was after the first trimester.

Acute puerperal inversion of the uterus in a primipara.

A rare case of uterine inversion immediately following delivery is reported. The uterus was successfully repositioned by a combined abdominal and vaginal approach.