The effects of dietary hesperidin supplementation on lamb performance and meat characteristics

An experiment was conducted to examine the effects of supplementing feed with hesperidin or α‐tocopheryl acetate on lambs' growth performance, carcass and meat characteristics. Sixteen male lambs were randomly assigned to four groups. One of the groups served as control and was given a basal diet, whereas the other three groups were given the same diet further supplemented with hesperidin at 1500 mg/kg, or hesperidin at 3000 mg/kg, or α‐tocopheryl acetate at 200 mg/kg. At the end of the experiment, lambs were fasted, weighed and slaughtered. After overnight chilling, samples of Longissimus thoracis muscle were taken and were used for meat quality evaluation. No significant differences were observed in final body weight, body weight gain and organ weights among the four groups. pH, color, water‐holding capacity, shear force values and intramuscular fat concentration of Longissimus thoracis muscle were also not significantly influenced by the dietary treatments. Measurement of lipid oxidation values showed that hesperidin supplementation positively influenced meat antioxidant properties during refrigerated storage.     

Seasonal change in the number of Cryptosporidium parvum oocysts in water samples from the rivers in Hokkaido,Japan, detected by the ferric sulfate flocculation method

An epidemiological study was carried out in natural water supplies of Hokkaido, one of the largest dairy prefectures in Japan. To investigate the prevalence of Cryptosporidium parvum (C. parvum) oocysts water samples were collected from three rivers in the eastern area of Hokkaido from August 1999 to October 2001, and C. parvum oocysts were collected and purified by the ferric sulfate flocculation method. The oocysts were detected using the immunofluorescent assay test (IFAT) and 4', 6-diamidino-2-phenylindole (DAPI) staining. The seasonal change in the number of oocysts detected was observed. Oocysts increased in numbers from the late summer to the early autumn (from August to November), thereafter, they exhibited a trend to decrease until December, when no oocysts could be detected. The maximum number of oocysts detected in the three rivers was 3.50, 5.00 and 3.33 oocysts/l, respectively. The oocyst density in river water changed in relation to the season in 1999, 2000 and 2001. This report first cleared up the seasonal changes in C. parvum oocysts number in river water.     

Viability and infectivity of Cryptosporidium parvum oocysts detected in river water in Hokkaido,Japan

The viability and infectivity of Cryptosporidium parvum (C. parvum) oocysts, detected in water samples collected from river water in Hokkaido, were investigated using Severe Combined Immunodeficient (SCID) mice. The water samples collected from September 27 through October 10, 2001 by filtration using Cuno cartridge filters were purified and concentrated by the discontinuous centrifugal flotation method. From 1.2 x 10 (5) liters of the raw river water, approximately 2 x 10(4) oocysts were obtained and designated as Hokkaido river water 1 isolate (HRW-1). Oocyst identification was carried out using microscopic and immunological methods. Six 8-week-old female SCID mice were each inoculated orally with 1 x 10 (3) oocysts. Infection was successfully induced, resulting in fecal oocyst shedding. Oocysts were then maintained by sub-inoculation into SCID mice every 3 months. Infectivity was evaluated by making comparisons with two known C. parvum stocks, HNJ-1 and TK-1, which were bovine genotypes detected in fecal samples from a cryptosporidiosis patient and young cattle raised in Tokachi, Hokkaido respectively. The oocyst genotypes were determined from a small subunit ribosomal RNA (SSU-rRNA) gene by polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) analysis. No significant differences were observed in the average number of oocysts per gram of feces (OPG) in any of the isolates. Our data indicates that the C. parvum oocysts detected in the sampled river water were of C. parvum genotype 2. Moreover, our data on the continued isolation, detection and identification of the C. parvum isolates is consistent with the available epidemiological data for the Tokachi area.     

Validation of a competitive ELISA for diagnosis of Brucella melitensis infection in sheep and goats

A competitive enzyme-linked immunosorbent assay (cELISA) was validated for the serodiagnosis of Brucella melitensis infection in small ruminants using 2108 positive and 2154 negative reference sera from sheep and goats. The optimum cut-off values, offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp), determined by receiver operating characteristic analysis, were at 23.6%, 21.8% and 25.0% inhibition of the conjugate control for sheep, goats and both species, respectively. The DSns of the cELISA for sheep, goats and both species at these cut-off values were 89.2% (95% confidence interval 87.1-91.1%), 74.0% (95% CI 71.4-76.5%) and 77.9% (95% CI 76.1-79.7%), whereas DSps were 96.4% (95% CI 95.2-97.4%), 92.9% (95% CI 91.1-94.3%) and 97.2% (95% CI 96.4-97.8%), respectively. Compared to cELISA, indirect ELISA and fluorescence polarisation assay have higher DSns and DSps. However, the results obtained with the cELISA were in good agreement with those of the complement fixation test (CFT) under field conditions using 5735 sheep and goat sera. The cELISA can be used as an alternative to the CFT for diagnosing B. melitensis infection in small ruminants.     

Sensitive and specific detection of Cryptosporidium species in PCR-negative samples by loop-mediated isothermal DNA amplification and confirmation of generated LAMP products by sequencing

Three LAMP (loop-mediated isothermal DNA amplification) assays were applied to detect Cryptosporidium species DNA in a total number of 270 fecal samples originating from cattle, sheep and horses in South Africa. DNA was extracted from 0.5 g of fecal material. Results of LAMP detection were compared to those obtained by nested PCR targeting the Cryptosporidium 18 small subunit rRNA (18S) gene. All samples were negative by nested PCR, while up to one-third of samples were positive by LAMP assays. The SAM-1 LAMP assay, shown to detect C. parvum, C. hominis and C. meleagridis, amplified Cryptosporidium DNA in 36 of 107 cattle (33.64%), in 26 of 85 sheep (30.5%) and in 17 of 78 horses (21.79%). The HSP LAMP specific to C. muris and C. andersoni, amplified Cryptosporidium DNA in one cow (0.9%), five sheep (5.8%) and seven horses (8.9%). The gp60 LAMP assay, shown to detect C. parvum produced no amplified Cryptosporidium DNA, likely due to low sample DNA concentrations. The specificity of LAMP assays was confirmed by sequencing of the LAMP products generated in positive samples. Sequence products from the three LAMP assays showed high identity to the target gene sequences confirming the specificity of LAMP. In this study, the LAMP procedure was clearly superior to nested PCR in the detection of Cryptosporidium species DNA. Use of LAMP is proposed as an efficient and effective tool for epidemiologic survey studies including screening of healthy animals in which Cryptosporidium oocyst shedding is characteristically low and likely below the detection limit of PCR in conventional sample concentrates.     

Serum liver enzyme activities in healthy Miniature Schnauzers with and without hypertriglyceridemia

OBJECTIVE: To determine whether hypertriglyceridemia in healthy Miniature Schnauzers is associated with high serum liver enzyme activities. DESIGN: Cross-sectional study. ANIMALS: 65 Miniature Schnauzers with serum triglyceride concentrations within the reference range (group 1), 20 Miniature Schnauzers with slightly high serum triglyceride concentrations (group 2), and 20 Miniature Schnauzers with moderately to severely high serum triglyceride concentrations (group 3). PROCEDURES: Questionnaires regarding each dog's medical history were completed, and serum alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and G-glutamyltransferase (GGT) activities were measured. RESULTS: Median serum ALP activity was significantly higher in group 3 than in group 1 or 2 dogs, but was not significantly higher in group 2 than in group 1 dogs. Median serum ALT activity was significantly higher in group 3 than in group 1 dogs, but was not significantly different between any of the other groups. Compared with group 1 dogs, group 2 and 3 dogs were significantly more likely to have high serum ALP activity (odds ratio, 26.2 and 192.6, respectively). Group 3 dogs also were significantly more likely to have high serum ALT activity (odds ratio, 8.0), serum AST activity (odds ratio, 3.7), and serum GGT activity (odds ratio, 11.3), compared with group 1 dogs. Group 3 dogs were significantly more likely (odds ratio, 31.0) to have > or = 2 high serum liver enzyme activities than were group 1 dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that moderate to severe hypertriglyceridemia was associated with high serum liver enzyme activities in Miniature Schnauzers.     

A study on ghrelin and LH secretion after short fasting and on ghrelin levels at perioestrual period in dairy cattle

In two experiments, we studied (a) the changes of LH secretion in heifers under different feeding schedules and (b) total ghrelin concentration at oestrus in cows and heifers. In experiment one, synchronized heifers were allocated in three groups (R, regularly fed controls; F, fasted; and F‐F fasted‐fed). One day after the completion of the oestrous induction protocol, group F and F‐F animals stayed without feed for 24 hr; thereafter, feed was provided to R and F‐F cattle; 2 hr later, GnRH was administered to all animals. Blood samples were collected for ghrelin, progesterone, LH and cortisol concentrations. Fasting caused increased ghrelin concentrations in groups F and F‐F, while in response to GnRH, LH surge was significantly attenuated in groups F and F‐F compared to R. In experiment 2, lactating cows and heifers were used. On day 9 of a synchronized cycle, PGF2α was administered, and blood samples were collected twice daily until the third day after oestrus and analysed for progesterone, estradiol, ghrelin, glucose and BHBA concentrations. No difference was recorded between groups in steroids and BHBA concentrations. In comparison to mid‐luteal values, ghrelin concentrations significantly increased at perioestrual period in cows, but not in heifers. This study provides evidence that starving‐induced elevated ghrelin concentrations can have suppressing effect on LH secretion, even after ghrelin's restoration to basal values and that during oestrus, ghrelin secretion is differently regulated in cows and heifers, likely being independent from oestradiol concentrations. Further research is required to identify the determining factors that drive the different regulation of ghrelin secretion in cows and heifers.     

ASSOCIATIONS BETWEEN DUAL‐PHASE COMPUTED TOMOGRAPHY FEATURES AND HISTOPATHOLOGIC DIAGNOSES IN 52 DOGS WITH HEPATIC OR SPLENIC MASSES

Ability to noninvasively differentiate malignant from nonmalignant abdominal masses would aid clinical decision making. The aim of this retrospective, cross‐sectional study was to identify features in dual‐phase computed tomographic (CT) studies that could be used to distinguish malignant from nonmalignant hepatic and splenic masses in dogs. Medical records were searched for dogs that had an abdominal dual‐phase CT examination, a hepatic or splenic mass, and subsequent histopathologic diagnosis. Computed tomographic images for all included dogs were acquired prior to and <30 s (early phase) and >60 s (delayed phase) after intravenous contrast administration. Fifty‐two dogs with 55 masses were studied: 24 hepatic, including 14 (58%) malignant and 10 (42%) non‐malignant; 31 splenic, including 18 (58%) malignant and 13 (42%) nonmalignant. There was substantial overlap in the pre‐ and postcontrast CT features of malignant and nonmalignant hepatic and splenic masses. Regardless of histologic diagnosis, hepatic masses most frequently showed marked, generalized enhancement in early phase images that persisted in the delayed phase. Splenic hemangiosarcoma and nodular hyperplastic lesions most frequently showed marked, generalized enhancement in early phase images that persisted in delayed images whereas most splenic hematomas had slight enhancement in early phase images. All splenic hematomas and 77% of the hemangiosarcomas had contrast accumulation compatible with active hemorrhage. There were no other significant differences in quantitative or categorical CT data between malignant and nonmalignant hepatic or splenic masses. Dual‐phase CT of dogs with hepatic or splenic masses provides limited specific diagnostic information.     

Investigation of hypertriglyceridemia in healthy Miniature Schnauzers

BACKGROUND: Idiopathic hypertriglyceridemia has been reported in Miniature Schnauzers (MS). However, studies investigating the prevalence of this disorder in a large population of MS are lacking. HYPOTHESIS: Hypertriglyceridemia is prevalent in healthy MS. ANIMALS: This study used 192 healthy MS and 38 healthy dogs of other breeds (control dogs). METHODS: Serum triglyceride and cholesterol concentrations were measured and statistically compared in both the MS and control group. Dogs were categorized based on their age, and median serum triglyceride concentrations were compared among different age groups. RESULTS: A total of 63 (32.8%) of the 192 MS had serum triglyceride concentrations above the reference range. In contrast, of the 38 control dogs, only 2 (5.3%) had serum triglyceride concentrations above the reference range. The median serum triglyceride concentration in MS was 73.5 mg/dL, which was significantly higher as compared to that of the control group (median, 55 mg/dL; P = .0005). Serum cholesterol concentration was above the reference range in 9 (9.0%) of 100 MS and in 2 (5.3%) of the control dogs. Mean serum cholesterol concentrations were not significantly different between the 2 groups (P = .1374). Median serum triglyceride concentrations in MS increased significantly with age (P < .0001), and there was a significant positive correlation between serum triglyceride concentrations and age (Spearman r = 0.47; P < .0001). There was no difference in serum triglyceride concentrations between male and female MS (P = .48). CONCLUSION: Healthy MS have a high prevalence of hypertriglyceridemia as compared to healthy dogs of other breeds. Both the prevalence and severity of hypertriglyceridemia increase with age.     

Response of ungrafted rootstocks and rootstocks grafted with wine grape varieties (Vitis sp.) to ‘lime-induced chlorosis’

The ungrafted rootstocks 41B, 1103P, 110R and 140Ru, the grafted combinations of 41B, 1103P and 110R with Xinomavro (one of the most important red wine grape varieties in Greece), as well as those of 1103P, 110R and 140Ru with Chardonnay, were evaluated for 'lime-induced chlorosis' tolerance by growing them with a) basic nutrient solution (BNS), b) BNS + 10 mM bicarbonate, c) BNS without iron (Fe) and d) BNS without zinc (Zn), in hydroponics. The ungrafted 140Ru followed by 41B under high bicarbonate presented the lowest degree of chlorosis; however only 41B presented non-differentiated biomass production and root/shoot ratio. Chlorotic symptoms in combination with plant growth parameters should be used as a tool for grapevine rootstock lime-tolerance screening whereas leaf Fe concentration and root ferric chelate reductase (FCR) activity should not. Lime-stress conditions affected plant mineral nutrition by depressing leaf nitrogen (N), phosphorus (P), calcium (Ca), magnesium (Mg) and increasing potassium (K), and zinc (Zn).