Molecular prevalence of Babesia bigemina in Rhipicephalus microplus ticks infesting cross-bred cattle of Punjab, India.

Babesiosis is an economically important tick-borne apicomplexan protozoan disease of cattle in tropical and subtropical regions. In the present study, Rhipicephalus microplus engorged female ticks were collected from 135 apparently healthy cattle from different agro-climatic zones of Punjab, India, to investigate the carrier status of Babesia bigemina infection in vector tick by using microscopy and PCR based assays. PCR when applied on DNA extracted from the egg masses harvested from ticks showed 1.48% (2/135) samples as positive, whereas 4.44% (6/135) samples were positive when product of primary PCR was used as template in nPCR. Further, among the DNA samples isolated from the unfed larval stages that emerged from egg masses laid by ticks, only 1.48% (2/135) samples were detected as positive for B. bigemina in PCR, while 7.41% (10/135) samples were detected positive in nPCR assay. Statistically, non-significant (p > 0.05) difference in prevalence rates was observed across different agro-climatic zones and between different age groups of cattle from which engorged ticks were collected. It can, thus, be concluded that prevalence of B. bigemina in the vector tick, R. microplus in Punjab state of India indicates an endemic status of the organism and a further study is needed for the management and control of the bovine babesiosis.

Molecular detection of Babesia bigemina infection in apparently healthy cattle of central plain zone of Punjab.

Bovine babesiosis is an important tick-borne disease caused by the parasites belonging to the genus Babesia, distributed worldwide and infecting a wide range of domesticated and wild cattle, occasionally man. The present study was conducted to determine the prevalence of B. bigemina infection in apparently healthy cattle from central plain zone of Punjab, India. Examination of Giemsa-stained peripheral thin blood smears revealed 2.45 % (5/204) animals to be positive for piroplasms of B. bigemina. However, genomic DNA isolated from these blood samples when subjected to primary PCR revealed a positivity of 7.35 % (15/204) as detected by the amplification of a 278-bp product in the agarose gel. PCR products obtained from the primary PCR of B. bigemina, when employed as template in nested PCR produced the amplicons of desired size (170 bp) was detected in 30.39 % (62/204) of the samples. It can thus be concluded that B. bigemina infection is prevalent in apparently healthy cattle population of this region and PCR assays can serve as a valuable tool for epidemiological studies in endemic areas.

Detection of acaricidal resistance in Hyalomma anatolicum anatolicum from Banaskantha district, Gujarat.

The resistance status in Hyalomma anatolicum anatolicum collected from Banaskantha district, Gujarat (India) was estimated by larval packet test (LPT) with different concentrations of amitraz (125, 250, 500, 750 and 1,000 ppm) and cypermethrin (100, 200, 300, 400 and 500 ppm). The regression graphs of mean mortality of larvae ticks were plotted against values of progressively increasing concentrations of amitraz and cypermethrin for the estimation of LC95 values and were determined as 1,529.39 and 351.84 ppm, respectively. Further a resistance of level I was determined against cypermethrin whereas, a comparatively higher resistance level (II) was recorded against amitraz. The current study appears to be the pioneer report of amitraz and cypermethrin resistance in H. a. anatolicum from the Gujarat state, India.

Prevalence of gastrointestinal helminth parasites of equids from organized farms of Mumbai and Pune.

A total of 1,304 faecal samples of different species of equids were examined for presence of helminthic eggs from five different centres in Mumbai and Pune regions of Maharashtra state from October 2011 to June 2012. Overall prevalence of helminthic infestation was found to be 20.63 % with higher rate of occurrence in monsoon (31.29 %) followed by winter (20.40 %) and summer (14.23 %). Four species of nematodes viz. strongyles (10.81 %), Strongyloides westeri (13.19 %), Parascaris equorum (0.23 %) and Dictyocaulus arnfieldi (0.23 %); two species of trematodes viz. amphistomes (1.38 %) and Schistosoma indicum (0.31 %) and only one species of cestode viz. Anoplocephala spp. (0.07 %) were encountered in the study. Coproculture studies revealed that Strongylus vulgaris was predominant species (36.87 %) among strongyles followed by cyathostomes. Amongst equids, mules had the lowest prevalence of 14.80 % followed by ponies (20.61 %) horses (38.79) and donkeys (51.90 %).

Prevalence of Theileria annulata infection in Hyalomma anatolicum anatolicum collected from crossbred cattle of Ludhiana, Punjab.

The prevalence of Theileria infection in tick vector Hyalomma anatolicum anatolicum collected from healthy animals of Ludhiana district, Punjab was recorded to assess the natural infection level of theilerial parasite in the field condition. A total of 60 semi-engorged H. a. anatolicum were collected from cattle and their salivary glands were dissected out. One half of the salivary gland was stained with methyl green pyronin (MGP) and the other half was utilized for DNA isolation for molecular detection of Theileria infection. A PCR and nested PCR assays were standardized for the detection of T. annulata infection in salivary gland of H. a. anatolicum. The prevalence of T. annulata infection was recorded as 8.3, 20.0 and 60.0 % by MGP staining, primary PCR and nested PCR, respectively. Further, the prevalence was higher in female ticks (8.8 %) than male ticks (6.6 %). The results demonstrated that both primary and nested PCR assays are a valuable technique for detection of T. annulata infection in vector tick under field conditions.

Comparison of nested PCR and microscopy for the detection of cryptosporidiosis in bovine calves.

The comparative efficacies of different conventional parasitological methods and nested PCR for diagnosis of bovine cryptosporidiosis in faecal samples were evaluated. Among the 100 samples collected from calves in and around Ludhiana Direct faecal smear staining technique revealed 25.0 % positivity for the oocysts of Cryptosporidium spp. with sensitivity and specificity of 68.12 and 92.98 %, respectively. Zinc sulphate solution floatation and saturated sugar solution floatation staining techniques showed sensitivity and specificity of 83.92 and 96.36; 81.03 and 98.14 %, respectively. Products of the primary PCR of Cryptosporidium spp. directed against small subunit (18S) ribosomal RNA when employed as template in nested PCR produced the amplicons of desired size (834 bp) in 47.0 % of the samples. Amplification of 834 bp fragment was also observed in positive control, while no amplification was observed in negative control. Results indicated PCR assays as highly sensitive and specific techniques for the screening of the samples for Cryptosporidium spp. but in developing countries and under field conditions where limited resources do not allow the application of PCR assays, concentration staining methods are recommended.

An in vitro biomechanical investigation of an interlocking nail system developed for buffalo tibia.

OBJECTIVE: The objectives of the study were to determine the mechanical properties of a customized buffalo interlocking nail (BIN), intact buffalo tibia, and ostectomized tibia stabilized with BIN in different configurations, as well as to assess the convenience of interlocking nailing in buffalo tibia. MATERIALS AND METHODS: The BIN (316L stainless steel, 12 mm diameter, 250 mm long, nine-hole solid nails with 10° proximal bend) alone was loaded in compression and three-point bending (n=4 each); intact tibiae and ostectomized tibiae (of buffaloes aged 5-8 years, weighing 300-350 kg) stabilized with BIN using 4.9 mm standard or modified locking bolts (4 or 8) in different configurations were subjected to axial compression, cranio-caudal three-point bending and torsion (n=4 each) using a universal testing machine. Mechanical parameters were determined from load-displacement curves and compared using Kruskal-Wallis test (p<0.05). RESULTS: Intact tibiae were significantly stronger than BIN and bone-BIN constructs in all testing modes. The strength of fixation constructs with eight locking bolts was significantly more than with four bolts. Overall strength of fixation with modified locking bolts was better than standard bolts. Based on technical ease and biomechanical properties, cranio-caudal insertion of bolts into the bone was found better than medio-lateral insertion. CLINICAL SIGNIFICANCE: The eight bolt BIN-bone constructs could be useful to treat tibial fractures in large ruminants, especially buffaloes.

Novel immunogenic baculovirus expressed virus-like particles of foot-and-mouth disease (FMD) virus protect guinea pigs against challenge.

Vaccination is a well accepted strategy for control of foot-and-mouth disease (FMD) in endemic countries. Currently, chemically inactivated virus antigens are used for preparation of FMD vaccine. To develop a non-infectious and safe recombinant vaccine, we expressed structural polypeptide of FMDV (O/IND/R2/75) using baculovirus expression system. We show that inclusion of mutated viral 3C protease in frame with the polypeptide (P1-2A), enhanced the yield of structural proteins. The structural proteins retained antigenicity and assembled into empty virus-like particles (VLPs). Immunization of guinea pigs with purified fractions of the VLPs resulted in humoral and cell mediated immune response by 4 weeks. The VLPs elicited comparable humoral immune response and relatively higher cell mediated immune response, when compared to conventional vaccine in guinea pigs. Further, up to 70% of the VLP immunized guinea pigs were protected against challenge with homologous guinea pig adapted virus. Our results highlight the application of recombinant FMDV VLPs in FMD vaccination.

Coprological investigation on neonatal bovine cryptosporidiosis in Ludhiana, Punjab.

An overall prevalence of 38.90 % was recorded for bovine cryptosporidiosis on coprological examination of 144 faecal samples collected from neonatal cattle calves from organised dairy farms in and around Ludhiana, Punjab by modified Ziehl-Neelsen staining. Further, a gradual decline in the percent prevalence was seen with increase in the age of the host from <1 month (64.1 %) to 4-5 months (12.5 %). The highest prevalence was recorded during the monsoon season (47.06 %) followed by summer (37.73 %) and winter (30.0 %) season and the seasonal variation was statistically significant (p ≤ 0.05). Female calves showed higher prevalence (44.32 %) than their male counterparts (27.66 %). Further, prevalence of cryptosporidiosis was significantly higher (p ≤ 0.05) in the diarrhoeic calves (52.70 %) as compared to the non-diarrhoeic (24.28 %) thus indicating a relatively higher risk (1.75 times) of the disease in diarrhoeic than normal calves.

[Retrospective analysis of one year of cesarean sections: indications and risk factors associated with intrapartum cesarean section]. / Analisi retrospettiva su un anno di tagli cesarei: indicazioni e fattori di rischio per taglio cesareo in travaglio di parto.

AIM: Cesarean section (CS) is currently the most commonly performed surgical intervention worldwide. Indications include previous CS, podalic presentation and fetal macrosomia in antepartum CS, fetal distress, and prolonged first or second phase of labor in intrapartum CS. Despite the marked reduction in fetal mortality and morbidity in selected circumstances, maternal complication rates associated with CS are far higher than with vaginal delivery. The aim of this study was to evaluate the indications and maternal and fetal outcomes in a population undergoing antepartum or intrapartum CS and to analyze the risk factors of intrapartum CS. METHODS: An analysis was conducted on data from 1748 deliveries performed at the Gynecology-Obstetrics Clinic of the University of Udine during 2006. A total of 603 medical records of the mothers who had delivered by CS and their infants were analyzed. The Indications and maternal and fetal outcomes were evaluated in mothers who had undergone elective CS, emergency antepartum or intrapartum CS; multivariate analysis was then performed to define the risk factors associated with intrapartum CS. RESULTS: Cesarean sections accounted for 34.5% of all deliveries performed during 2006, of which 42.1% were elective CS, 21.1% emergency antepartum CS, and 36.8% intrapartum CS. The principal reason for CS delivery was previous CS in elective CS, gestational hypertension in emergency antepartum CS, and cardiotocographic alterations in emergency intrapartum CS. Multivariate logistic regression analysis showed that the independent risk factors for intrapartum CS were excessive weight gain during pregnancy and medically induced labor. CONCLUSION: A more accurate evaluation of the mother's and neonate's needs could help to improve the current rates of CS. This could be achieved by selection of cases in which there is a real need for CS and by acting on modifiable risk factors of intrapartum CS such as excessive weight gain during pregnancy.

Novel antibodies in the treatment of non-Hodgkin's lymphoma.

Monoclonal antibodies (mAbs) have revolutionised the treatment of malignancies, especially non-Hodgkin's lymphoma (NHL). Antibody-based therapies target tumour cells expressing a specific antigen while sparing the majority of normal cells leading to a decrease in treatment-associated toxicity. Rituximab, a monoclonal antibody directed against CD20 on B cells, was the first monoclonal antibody to be approved by the US Food and Drug Association (FDA) in 1997 for the treatment of patients with relapsed/refractory, follicular or low-grade NHL . However, it was soon realised that not all patients respond to rituximab therapy and close to 60% of patients with follicular lymphoma who were previously sensitive to rituximab become 'resistant' to repeat rituximab therapy. This led to further attempts to improve the antitumour activity of anti-CD20 mAbs (i.e. 2nd/3rd generation anti-CD20s), and to identify additional potential targets on lymphoma cells other than CD20. A number of these antibodies directed against lymphoma cell targets other than CD20 are now undergoing development, many of which are currently in clinical trials. This manuscript focuses on an overview of these 'non-anti-CD20' novel mAbs for NHL.