Host transcriptome response to heat stress and Eimeria maxima infection in meat-type chickens.

Eimeria (E.) maxima parasite infects chickens' midgut disrupting the jejunal and ileal mucosa causing high morbidity and mortality. Heat stress (HS) is a seasonal stressor that impacts biological functions leading to poor performance. This study elucidates how HS, E. maxima infection, and their combination affect the ileum transcriptome. Two-hundred and forty 2-week-old males Ross708 chickens were randomly allocated into four treatment groups: thermoneutral-control (TNc), thermoneutral-infected (TNi), heat-stress control (HSc), and heat stress-infected (HSi), with 6 replicates each of 10 birds. Infected groups received 200x103 sporulated E. maxima oocysts/bird, and heat-treated groups were raised at 35°C. At 6-day post-treatment, ileums of five randomly selected chickens per group were sampled, RNA was extracted and sequenced. A total of 413, 3377, 1908, and 2304 DEGs were identified when applying the comparisons: TNc vs HSc, TNc vs TNi, HSi vs HSc, and TNi vs HSi, respectively, at cutoff ≥1.2-fold change (FDR: q<0.05). HSc vs TNc showed upregulation of lipid metabolic pathways and degradation/metabolism of multiple amino acids; and downregulation of most immune-related and protein synthesis pathways. TNc vs TNi displayed upregulation of most of immune-associated pathways and eukaryotic mRNA maturation pathways; and downregulation of fatty acid metabolism and multiple amino acid metabolism pathways including tryptophan. Comparing HSi versus HSc and TNi revealed that combining the two stressors restored the expression of some cellular functions, e.g., oxidative phosphorylation and protein synthesis; and downregulate immune response pathways associated with E. maxima infection. During E. maxima infection under HS the calcium signaling pathway was downregulated, including genes responsible for increasing the cytoplasmic calcium concentration; and tryptophan metabolism was upregulated, including genes that contribute to catabolizing tryptophan through serotonin and indole pathways; which might result in reducing the cytoplasmic pool of nutrients and calcium available for the parasite to scavenge and consequently might affect the parasite's reproductive ability.

Effect of Glucose Supplementation on Apoptosis in the Pectoralis major of Chickens Raised under Thermoneutral or Heat Stress Environment.

Reduced feed intake during heat stress (HS) disrupts glucose homeostasis, thereby resulting in endoplasmic reticulum (ER) stress and triggering apoptosis in chickens. We hypothesize that glucose supplementation could reduce apoptosis in chickens raised under HS. This study comprised 456 28-day-old broiler chickens randomly assigned to four treatment combinations under glucose supplementation and HS. The treatments were TN0, TN6, HS0, and HS6 with two glucose levels (0% and 6%) and two temperature levels (25 °C (thermoneutral-TN) and 35 °C (8.00 AM to 8.00 PM, (HS)). After 7 days post-HS, the blood glucose level for the HS6 group was higher than for TN0, TN6, and HS0. We studied the mRNA expression of genes and caspase-3 activity in the four experimental groups. The expressions of GCN2, ATF4, CHOP, and FOXO3a increased during HS regardless of glucose supplementation, while PERK and MAFbx increased only under HS with glucose supplementation. We show that under TN conditions, glucose supplementation led to a significant increase in cellular apoptosis in the Pectoralis (P.) major. However, under HS with glucose, the level of apoptosis was similar to that of chickens raised under TN conditions with no glucose supplementation. The utility of glucose to curtail apoptosis under HS should be tested under other intense models of HS.

Glucose Supplementation Improves Performance and Alters Glucose Transporters' Expression in Pectoralis major of Heat-Stressed Chickens.

Glucose level in birds' tissue decreases due to heat stress (HS)-induced reduction in feed intake (FI); impairing metabolism and growth. The effect of glucose supplementation on the performance of broiler chickens was evaluated under thermoneutral (TN) and HS conditions. Glucose was supplemented at 0 and 6% under TN-(25 °C) and HS-(25 °C-35 °C-25 °C) conditions. The treatments were TN + 0%-glucose (TN0); TN + 6%-glucose (TN6), HS + 0%-glucose (HS0) and HS + 6%-glucose (HS6). There were 6 replicates (19 birds each)/treatment. Heat and glucose supplementation were applied from d28-35. At d35, Pectoralis (P.) major was sampled from one bird/replicate to determine glucose transporters' mRNA expression. Heat application lowered (p < 0.05) FI, body weight gain, and increased feed and water conversion ratios. Glucose supplementation increased total energy intake by 4.9 and 3.2% in TN and HS groups, respectively but reduced FI under TN and HS conditions. The P. major- and drumstick-yield reduced (p < 0.05) in HS0 compared to TN0, TN6 and HS6. Under HS, glucose supplementation improved eviscerated carcass weight by 9% and P. major yield by 14%. Glucose supplementation increased SGLT1 expression with/without heat treatment while HS independently increased the expression of GLUT 1, 5 and 10. Glucose supplementation under HS could improve performance of broilers.

Quantitative and molecular aspects of water intake in meat-type chickens.

Even though water is the most essential nutrient for poultry production, adequate data on individual water intake in broiler chickens and its relationship with other traits of economic importance is scant. Water is provided to chickens in an unrestricted manner in spite of being a finite resource. Climate change continues to affect water sources and efficient bird use of water is long overdue. Understanding the biological basis of water intake is essential for sustainability of the poultry industry. Individual water and feed intake, and growth data was collected on 520 commercial broilers aged 14 to 42 days. We introduced the concepts of water conversion ratio (WCR) and residual water intake (RWI) as parameters that can be used to assess water intake efficiency. Water conversion ratio was defined as the amount of water consumed per unit of body weight gain, and RWI was defined as the difference between the actual water intake (WI) of a given bird and the expected WI by an average bird from the population with the same metabolic body weight, feed intake (FI) and body weight gain (BWG). The correlation between WI and FI was positive (r=0.77; P<0.0001), and the correlation between WI and BWG was positive (r=0.80; P<0.0001). Based on the distribution of RWI, the bottom 5 birds (LRWI) and the top 5 birds (HRWI) for RWI were selected for mRNA expression differences. The average broiler consumed about 7.8 L (± 1L) of water from 14 to 42 days of age. The mRNA expression of arginine vasopressin (AVP) antidiuretic hormone, calcium sensing receptor (CasR), sodium channel epithelial 1 subunit alpha (SCNN1A) and SCNN1D in the hypothalamus was upregulated in the LRWI group compared to the HRWI group. Similarly, kidney aquaporins (AQP) 2, 3, and 4 were upregulated in the LRWI group compared with the HRWI group. Given that water was provided ad libitum, the up-regulation of AVP and AQP gene mRNA expressions seem to indicate that the LRWI birds were more efficient in water reabsorption in the kidney compared to their HRWI counterparts. Increased water reabsorption will reduce the amount of water consumed to attain hydration. The water reabsorption potential was reflected in the excreta moisture levels as the LRWI birds had significantly lower excreta moisture than the HRWI birds. Excreta moisture level require further studies and could be considered as a potential proxy trait for water intake.

Selective determination of nitrite in water and food samples using zirconium oxide (ZrO2)@MWCNTs modified screen printed electrode.

Nitrite ions are being used in different forms as food preservatives acting as flavor enhancers or coloring agents for food products. However, continuous ingestion of nitrite may have severe health implications due to its mutagenic and carcinogenic effects. Thus, this study constructed an electrochemical assay using disposable nano-sensor chip ZrO2@MWCNTs screen printed electrodes (SPE) for the rapid, selective, and sensitive determination of nitrite in food and water samples. As a sensing platform, the use of nanomaterials, including metal oxide nanostructures and carbon nanotubes, exhibited a superior electrocatalytic activity and conductivity. Morphological, structural, and electrochemical analyses were performed using electron microscopy (SEM and TEM), Fourier-transform infrared (FTIR) spectroscopy, electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and chronoamperometry (CA). Accordingly, a wide dynamic linear range (5.0 µM to 100 µM) was obtained with a limit of detection of 0.94 µM by the chronoamperometric technique. In addition, the sensor's selectivity was tested when several non-target species were exposed to the sensor chips while no obvious electrochemical signals were generated when the nitrite ions were not present. Eventually, real food and water sample analysis was conducted, and a high recovery was achieved.

Temporal dynamics of the cecal and litter microbiome of chickens raised in two separate broiler houses.

In this study, we investigated the dynamics of the ceca and litter microbiome of chickens from post-hatch through pre-harvest. To achieve this, six hundred one-day old Cobb 500 broiler chicks were raised on floor pens for 49 days in two separate houses. We performed short-read and full-length sequencing of the bacterial 16S rRNA gene present in the meconium and in cecal and litter samples collected over the duration of the study. In addition, we determined the antimicrobial resistance (AMR) phenotype of Escherichia coli and Enterococcus spp. isolated from the meconium and the ceca of 49-day old chickens. We monitored the relative humidity, temperature, and ammonia in each house daily and the pH and moisture of litter samples weekly. The overall microbial community structure of the ceca and litter consistently changed throughout the course of the grow-out and correlated with some of the environmental parameters measured (p < 0.05). We found that the ceca and litter microbiome were similar in the two houses at the beginning of the experiment, but over time, the microbial community separated and differed between the houses. When we compared the environmental parameters in the two houses, we found no significant differences in the first half of the growth cycle (day 0-21), but morning temperature, morning humidity, and ammonia significantly differed (p < 0.05) between the two houses from day 22-49. Lastly, the prevalence of AMR in cecal E. coli isolates differed from meconium isolates (p < 0.001), while the AMR phenotype of cecal Enterococcus isolates differed between houses (p < 0.05).

Application of Amniotic Membrane in Skin Regeneration.

Amniotic membrane (AM) is an avascular structure composed of three different layers, which contain collagen, extracellular matrix, and biologically active cells (stem cells). Collagen, a naturally occurring matrix polymer, provides the structural matrix/strength of the amniotic membrane. Tissue remodeling is regulated by growth factors, cytokines, chemokines, and other regulatory molecules produced by endogenous cells within AM. Therefore, AM is considered an attractive skin-regenerating agent. This review discusses the application of AM in skin regeneration, including its preparation for application to the skin and its mechanisms of therapeutic healing in the skin. This review involved collecting research articles that have been published in several databases, including Google Scholar, PubMed, Science Direct, and Scopus. The search was conducted by using the keywords 'amniotic membrane skin', 'amniotic membrane wound healing', 'amniotic membrane burn', 'amniotic membrane urethral defects', 'amniotic membrane junctional epidermolysis bullosa', and 'amniotic membrane calciphylaxis'. Ultimately, 87 articles are discussed in this review. Overall, AM has various activities that help in the regeneration and repair of damaged skin.

Mannose-methyl-ß-cyclodextrin suppresses tumor growth by targeting both colon cancer cells and tumor-associated macrophages.

Methylated ß-cyclodextrin (MßCD) can extract cholesterol from lipid rafts and induce apoptosis in cancer cells by inhibiting activation of the PI3K-Akt-Bad pathway. In this study, we modified MßCD with mannose (Man-MßCD) and assessed its in vitro and in vivo potential for targeting colon cancer cells expressing the mannose receptor (MR) and tumor-associated macrophages (TAM). Man-MßCD showed a significantly greater level of cellular association with colon-26 cells and M2 macrophages, and much more prominent anticancer activity than that of MßCD against MR-positive colon-26 cells. These results revealed that autophagy was the main mechanism of cell death associated with Man-MßCD. Furthermore, compared with MßCD, Man-MßCD significantly reduced tumor development following intravenous delivery to tumor-bearing mice, with no apparent side effects. Thus, Man-MßCD has the potential to be a novel anticancer drug.

Heat Stress Alters the Effect of Eimeria maxima Infection on Ileal Amino Acids Digestibility and Transporters Expression in Meat-Type Chickens.

Eimeria (E.) maxima invades the midgut of chickens and destroys the intestinal mucosa, impacting nutrient digestibility and absorption. Heat stress (HS) commonly affects the broiler chicken and contributes to inflammation and oxidative stress. We examined the independent and combined effects of HS and E. maxima infection on apparent amino acid ileal digestibility (AID) and mRNA expression of amino acid transporters in broiler chickens (Ross 708). There were four treatment groups: thermoneutral-control (TNc) and infected (TNi), heat-stress control (HSc) and infected (HSi), six replicates of 10 birds/treatment. Ileal content and tissue were sampled at 6 d post infection to determine AID and transporters expression. Surprisingly, the HSi chickens exposed to two critical stressors exhibited normal AID. Only the TNi group displayed reduction in AID. Using TNc as control, the HSc group showed upregulated CAT1, LAT4, TAT1, SNAT1, and SNAT7. The HSi group showed upregulated CAT1 and LAT1, and downregulated b0,+AT, rBAT, SNAT1, and SNAT2. The TNi group showed upregulated CAT1, LAT1, and SNAT1 and downregulated B0AT1, b0,+AT, rBAT, LAT4, and TAT1. The expression of all enterocytic-apical and about half of the basolateral transporters was higher in the HSi group than in the TNi group, indicating that HS can putatively alleviate the E. maxima adverse effect on ileal digestion and absorption.

Heat stress modulates the disruptive effects of Eimeria maxima infection on the ileum nutrient digestibility, molecular transporters, and tissue morphology in meat-type chickens.

Eimeria (E.) maxima is one of the most pathogenic Eimeria spp persistently invading the middle jejunum and ileum, damaging the intestinal mucosa of chickens. Heat stress (HS) is a common stressor and equally contributes to inflammation and oxidative stress. We investigated the effect of E. maxima infection and HS on ileal digestibility, mRNA expression of nutrient transporters, and ileal tissue morphology in broiler chickens. There were four treatment groups: thermoneutral control (TNc), thermoneutral infected (TNi), heat stress control (HSc), and heat stress infected (HSi), 6 replicates each of 10 birds per treatment. Chickens were fed a diet containing 0.2% TiO2. At 6-day-post infection, ileal content and tissue were collected to quantify ileal digestibility of crude protein and fat, mRNA levels of nutrient transporters and histopathology. Growth and feed intake were reduced in all treatment groups, compared with the TNc. Contrary to expectation, the combination of two major stressors (E. maxima and HS) in the TNi group exhibited almost normal digestibility while only the TNi birds expressed severe digestibility depression, compared with the TNc group. The TNi group showed the lowest mRNA expression of the transporters: SGLT1, GLUT2-5-8-10-12, FABP1-2-6, and PEPT1 compared with the other treatment groups. The expression of the absorptive enterocytes' gene markers (ACSL5, IAP, and SGLT1) supported by the ileal tissue morphology indicated that the TNi group had the highest enterocytic destruction. The expression of oxidative genes (iNOS and CYBB) dramatically increased only in the TNi group compared with the other treatment groups. Our results showed that exposing broiler chickens to HS can mitigate the disruptive effect of E. maxima on the ileal digestibility and absorption by limiting the parasite-induced tissue injury and suppressing the enterocytic inducible oxidative damage.

A Study on Outcome of Surgical Treatment of Chronic Retrocalcaneal Bursitis.

Retrocalcaneal bursitis is one of the important causes of posterior heel pain which is due to repetitive friction of the retrocalcaneal bursa between the postero-superior calcaneal tuberosity (haglund deformity) and Achilles tendon. Most of the patients are treated by conservative methods. But when the condition becomes chronic and not responding to the conservative treatment, surgical treatment is an option of these cases. This prospective interventional study was conducted in the Department of Orthopaedic Surgery, BSMMU (Bangabandhu Sheikh Mujib Medical University), Dhaka, Bangladesh from September 2017 to August 2019. Within this period, total 40 cases of chronic retrocalcaneal bursitis were operated at BSMMU. The clinical and functional outcome was evaluated according to AOFAS (American Orthopedic Foot Ankle Society) scale. The results of this study showed significantly improvement of pain, functional status and deformity at the time of final follow-up period of 1 year, total mean score improved from 46/100 to 89/100. The outcome of the subjects was satisfactory 90% and unsatisfactory 10%. Surgical intervention is an effective option of treatment with satisfactory outcome, in the treatment of chronic retrocalcaneal bursitis.

Association of miR-146a rs57095329 with Behçet's disease and its complications.

BACKGROUND: Behçet's disease is a chronic relapsing and remitting autoimmune multisystem inflammatory disease characterised by oral aphthae, genital ulcers, skin lesions, gastrointestinal involvement, arthritis, vascular lesions and neurological manifestations. We hypothesised a link between rs57095329 of miR-146a and Behçet's disease, with further links with common clinical features. METHODS: We tested our hypothesis in 130 Behçet's disease patients and 131 age and sex-matched healthy controls. Behcet's disease current activity index (BDCAI) was used to assess patients' disease activity status. MiR-146a (rs57095329) was genotyped in all participants using RT-PCR and results in patients analysed according to clinical features. RESULTS: The frequency of the GG and AG genotypes in rs57095329 were strongly associated with Behçet's disease (adjusted OR 8.05, 95% CI 3.63-17.82; P < 0.001 and OR 2.26, 95% CI 1.27-4.04; P = 0.006, respectively), and in dominant (GG+AG > AA) and recessive (GG > AA+AG) models (both P < 0.001). Additionally, G allele distribution was significantly greater in Behçet's disease compared with controls (OR 2.85, 95% CI 1.98-4.11, P < 0.001). The AA genotype and A allele were linked to oral ulcers, the GG genotype and G allele to neurological disease, and the GG genotype and G allele to ocular disease (all P < 0.01). There were no links with genital ulceration, skin lesions, vascular disease or the result of the pathergy test. CONCLUSION: The miR-146a (rs57095329) is associated with Behçet's disease and certain genotypes and alleles with oral ulcers, and with ocular and neurological manifestations.

Heat stress reduces sexual development and affects pathogenesis of Eimeria maxima in meat-type chickens.

Coccidiosis, caused by Eimeria spp. presents a self-limiting intestinal infection of poultry. Intestinal replication of the parasite causes severe morphological alterations to the host gastrointestinal tract, marked, among others, by the disruption of the intestinal barrier. We have previously reported a significant reduction in merozoite replication and oocyst shedding in E. tenella in vitro and in vivo. The objective of this study was to investigate the pathogenesis of E. maxima infection in broiler chickens under heat stress (HS) and mRNA expression of host cytokines that might affect the curtailed development of the parasite. We herein demonstrate that there is a significant detrimental effect of HS on the pathogenesis of E. maxima infection in broilers. There was a restricted replication of the parasite in HS chickens evidenced by significantly reduced oocyst shedding and disruption of the intestinal blood barrier. Gene expression of parasite genes demonstrated curtailed sexual reproduction of E. maxima in HS chickens. There was downregulation of Eimeria spp. genes related to gamete fusion, oocyst shedding, mitosis and spermiogenesis. Host gene expression indicates alterations in the cytokine expression that could be related to reduced parasite development in vivo.

Molecular diagnosis and clinico-hemato-biochemical alterations and oxidant-antioxidant biomarkers in Babesia-infected dogs of Mizoram, India.

BACKGROUND & OBJECTIVES: Babesiosis is a tick transmitted disease, infecting a wide variety of wild and domestic animals, as well as humans. This study was designed to investigate molecular diagnosis and clinic-hemato-biochemical and oxidant/antioxidant status in dogs of Mizoram, India. METHODS: A total 1200 dogs screened for babesiosis during 2017-18 and 53 dogs suspected for babesiosis by clinical signs and were confirmed by molecular diagnosis. Clinical signs were recorded; also blood samples were taken to investigate hematologic changes, serum biochemical variations and oxidative stress biomarkers. RESULTS: The overall incidence of babesiosis in dogs of Aizawl, Mizoram, India during the study period recorded was 1.25% (15/1200) and 28.3% cases confirmed from 53 suspected dogs (15/53). The most commonly observed clinical signs were fever, emaciation, depression and icterus and lymphadenopathy. Significant reduction in PCV, HB, RBCs, MCHC, total protein, and albumin along with significant increase in MCV, WBCs, monocytes and BUN were the most consistent hemato-biochemical changes. Oxidant/antioxidant assessment showed significant reduction in superoxide dismutase, catalase and total anti-oxidant (TAC) along with significant increase in lipid peroxidase (LPO) activities. INTERPRETATION & CONCLUSION: The findings of this study demonstrated that the main causative agent of babesiosis in dogs in Mizoram Province is Babesia gibsoni which caused significant alteration of hemato-biochemical and oxidant-antioxidant status in dogs.

Pattern of induced estrus and conception rate following Ovsynch and Ovsynch based gonadotropin-releasing hormone treatments initiated on day 6 of estrous cycle in repeat breeding crossbred cows.

AIM: The aim was to evaluate the estrus response, incidence of accessory corpus luteum formation and fertility following different hormonal protocols in repeat breeding crossbred cows. MATERIALS AND METHODS: This study was carried out on 24 repeat breeding crossbred cows allotted into four groups. Cows of Group I was not given any treatment, Group II was treated with gonadotropin-releasing hormone (GnRH) injection on day 6 post-estrus, Group III was treated with Ovsynch protocol, and Group IV was treated with Ovsynch based GnRH treatment. Estrus responses such as duration, onset, percentage, and intensity of estrus were recorded during the study. The incidence of accessory corpus luteum was recorded per rectally on day 7 after first and additional GnRH of Ovsynch treatment. The conception rate for all groups was calculated by the absence of estrus and on day 45 after artificial insemination (AI) per rectum. Serum samples were collected at AI and day 12 post-AI in Group I and II. Serum samples were also collected at GnRH, Prostaglandin F2α (PGF2α), timed AI (TAI) and day 12 post-TAI in Group III and IV. RESULTS: Ovsynch and Ovsynch based GnRH treatments are resulted in 100.00% induction of estrus after the PGF2α injection. Onset of induced estrus after the PGF2α injection for Group III and IV was recorded as 48.750±0.713 and 51.472±1.989 h, respectively, and it was not significant. There was no significant difference in duration of estrus among the groups. The incidence of intermediate estrus intensity was found to be highest. All the cows showed the incidence of formation of accessory corpus luteum subsequent to GnRH treatment on day 6 of the estrous cycle in Group II, III, and IV. The conception rate was 0.00%, 16.67%, 50.00%, and 50.00% in Group I, II, III, and IV, respectively. CONCLUSION: Ovsynch and Ovsynch based GnRH treatments initiated on day 6 of estrous cycle capable of responding with a higher percentage of ovulation and formation of accessory corpus luteum which helped in higher conception rate over single post-AI GnRH treatment in repeat breeders. These treatments responded with better estrus response but did not significantly improve estrus intensity.

Anti-inflammatory activities of some newly synthesized pyridinyl- and indazolyl benzamide derivatives.

A series of substituted (pyridin-4-yl)phenyl-2-methoxybenzamide and their derivatives were prepared and screened for their anti-inflammatory activities. Initially the acute toxicity of the compounds was assayed via the determination of their LD50. Some of the newly synthesized compounds exhibited better pharmacological and biological responses than the reference controls with low concentrations. The structures of newly synthesized compounds were confirmed by chemical, elemental and spectroscopic evidences.

Correlation between peritoneal lavage cytology and tumour stage in patients with colorectal cancer.

BACKGROUND AND STUDY AIMS: Complete surgical removal of the involved bowel segment in colorectal cancer is the most effective primary treatment. The main prognostic factors for colorectal cancer are penetration of the tumour into different layers of the bowel wall and regional lymph node involvement. Positive lavage cytology has been used to predict peritoneal recurrence, but its effectiveness remains controversial. This study was conducted to assess the prevalence of positive peritoneal lavage cytology in correlation with the tumour stage in patients with colorectal cancer. PATIENTS AND METHODS: This prospective cross-sectional study was performed on 20 patients with different cases of colorectal cancer attending the colorectal unit and emergency department of the Kasr Al Ainy Hospital, Cairo University Hospitals, from March 2012 to March 2013. RESULTS: The patients' gender did not influence the peritoneal lavage cytology results (p = 0.062); there is no significant correlation between the TNM staging system and cytology in patients with colorectal cancer (p = 0.253). CONCLUSION: Although there is a positive linear correlation between the tumour stage and positive peritoneal lavage cytology, it did not reach a statistically significant level. In addition, the greater the depth of invasion, the higher the lavage cytology rate. However, this trend was not statistically significant.

Characterization of shiga toxin producing (STEC) and enteropathogenic Escherichia coli (EPEC) in raw yak (Poephagus grunniens) milk and milk products.

Thirty-one shiga toxin-producing (STEC) and 6 enteropathogenic Escherichia coli (EPEC) were isolated from 87 raw yak milk and 63 'churpi' samples. Of 18 stx(1) positive isolates (48.6%), 14 carried stx(1c) (77.7%). Subtyping of 28 stx(2) positive isolates (75.7%) revealed the presence of stx(2c) (9, 32.1%), stx(2d) (3, 10.7%), stx(2e) (1, 3.57%) and stx(2f) (3, 10.7%) variants. Furthermore, intimin (eaeA), enterohaemolysin (ehxA), autoagglutinating adhesin (saa), iha (adherence conferring protein), efa1 (EHEC factor for adherence), bundle forming pilli (bfpA) and toxB (type III secreted protein encoded on LEE Island, similar to toxin B of Clostridium difficile) genes were detected in 14, 16, 12, 4, 3, 2 and 2 isolates, respectively. Univariate and multivariate analysis depicted that both stx(1) and stx(2) or their variants were more likely to occur in isolates from Arunachal Pradesh (p<0.04) rather than Sikkim. Dendogram constructed on the basis of RAPD and ERIC PCR profile distributed the STEC and EPEC isolates in separate clusters irrespective of their sources and serotypes. The STEC and EPEC isolates exhibited resistance against erythromycin, amikacin, azithromycin, amoxicillin, ampicillin+cloxacillin, cephalothin, furazolidone, gentamicin, kanamycin, streptomycin and tetracycline. This is the first ever report on occurrence and characterization of STEC and EPEC isolated from yak milk and milk products.

Efficacy of Heatsynch protocol for induction of estrus, synchronization of ovulation and timed artificial insemination in yaks (Poephagus grunniens L.).

The objective of this study was to test the efficacy of induction of estrus, synchronization of ovulation and timed artificial insemination in anestrous yaks using the Heatsynch protocol. In Experiment 1, 10 anestrous yaks were administered an analogue of gonadotropin releasing hormone (GnRH) followed by prostaglandin (PG)F2alpha 7 days later and then estradiol cyponate (ECP) 24 h after that. Ovulation was detected by rectal palpation at 2h intervals beginning at the initial signs of estrus. Blood samples were collected at 2h intervals beginning at the time of ECP injection up to 2h after the occurrence of ovulation for the determination of LH and progesterone. All the animals responded to the Heatsynch protocol with expression of estrus and synchronization of ovulation. The mean time interval from the ECP injection to ovulation was 59.4+/-2.62 h (range 50-72 h). The interval from the LH peak to ovulation was 30.2+/-2.3 h. The high degree of synchrony in ovulation could be attributed to the synchrony in the timing of LH peaks. In Experiment 2, 10 anestrous yaks were treated with the Heatsynch protocol (as in Experiment 1) and TAI was performed at 48 and 60 h after the ECP treatment. Concurrently, 16 cycling yaks were inseminated approximately 12 h after detection of spontaneous estrus. Pregnancy rates were similar in both groups, 40% for TAI and 43.75% for yaks inseminated following spontaneous estrus (p>0.05). From this study, two conclusions can be drawn. First, the Heatsynch protocol can be successfully used to induce and synchronize estrus in anestrous yaks and, second, ovulation following the Heatsynch protocol is synchronized adequately to permit the use of fixed time AI in this species.

Potentiometric batch and flow injection analysis of betaine hydrochloride.

Novel PVC membrane electrodes for the determination of betaine ion based on the formation of betaine-tetraphenylborate (Be-TPB) and betaine-phosphotungstate (Be-PT) ion-exchangers as electroactive materials are described. The sensors show a fast, stable, near Nernstian response for 6.92 x 10(-6) to 7.94 x 10(-3) M and 1.0 x 10(-4) to 1.0 x 10(-2) M betaine hydrochloride (Be.Cl) in case of Be-TPB electrode applying batch and flow injection analysis (FIA), respectively, and 2.95 x 10(-5) to 2.26 x 10(-3) M and 3.16 x 10(-5) to 1.0 x 10(-2) M in case of Be-PT electrode for batch and FIA electrodes, respectively, at 25 degrees C over the pH range of 3.5-10 with a cationic slope of 60.2 and 59.1 mV decade(-1) and a fast potential response of < or =15 s. The lower detection limits are 7.94 x 10(-6) and 3.18 x 10(-5) M Be.Cl for Be-TPB and Be-PT electrodes, respectively. Selectivity coefficient data for some common inorganic cations, sugars, amino acids and the components other than betaine, of the mixed drug investigated show negligible interference. The electrodes have been applied to the direct potentiometric determination of betaine hydrochloride in water and in a pharmaceutical preparation under batch and FIA conditions. Potentiometric titrations of Be.Cl with NaTPB and PTA as titrants were monitored with the developed betaine electrodes as an end point indicator electrode. The determination of Be.Cl shows an average recovery of 100.8% with mean relative standard deviation of 0.61%. The effect of temperature on the electrodes was also studied.