Antioxidative Effects of Astaxanthin against Nitric Oxide‐Induced Oxidative Stress on Cell Viability and Gene Expression in Bovine Oviduct Epithelial Cell and the Developmental Competence of Bovine IVM/IVF Embryos

The aim of the present study was to elucidate the fundamental mechanism of bovine oviduct epithelial cell (BOEC) co‐culture on developmental capacity of bovine in vitro oocyte maturation/in vitro fertilization (IVM/IVF) embryos. We examined the effects of astaxanthin against nitric oxide‐induced oxidative stress on cell viability by MTT assay, lipid peroxidation (LPO) by using thiobarbituric acid (TBA) reaction for malondialdehyde (MDA) and the expression of antioxidant genes (CuZnSOD, MnSOD and Catalase) or apoptosis genes (Bcl‐2, Caspase‐3 and Bax) by RT‐PCR in BOEC. We also evaluated the developmental rates of bovine IVM/IVF embryos co‐cultured with BOEC pre‐treated with astaxanthin (500 μm ) in the presence or absence of sodium nitroprusside (SNP, 1000 μm ) for 24 h. Cell viability in BOEC treated with SNP (50–2000 μm ) lowered, while astaxanthin addition (50–500 μm ) increased it in a dose‐dependent manner. Cell viability in astaxanthin plus SNP (1000 μm ) gradually recovered according to the increase in astaxanthin additions (100–500 mm ). The LPO in astaxanthin group (50–500 μM) gradually decreased in a dose dependent manner and among SNP or astaxanthin plus SNP group, SNP alone and astaxanthin (50 μM) plus SNP shown a significant increase than other groups (p < 0.05). Expression of apoptosis or antioxidant genes was detected by RT‐PCR. Bcl‐2 and antioxidant genes were detected in astaxanthin or astaxanthin plus SNP group, and Caspase‐3 and Bax genes were only found in SNP group. When bovine IVM/IVF embryos were cultured for 6–7 days under co‐culture system such as BOEC treated with astaxanthin in the presence or absence of SNP, the developmental ability to blastocysts in 500 μm astaxanthin group was the highest of all groups. These results suggest that astaxanthin has a antioxidative effect on cell viability and LPO of BOEC, and development of bovine IVM/IVF embryos due to the induction of antioxidant genes and suppression of apoptosis genes.     

Probability of and risk factors for introduction of infectious diseases into Dutch SPF dairy farms: a cohort study

A 2-year cohort study was conducted to investigate the probability of disease introduction into Dutch dairy farms. The farms were tested regularly for diseases and were visited biannually to collect management data. Ninety-five specific pathogen-free (SPF) dairy farms were selected from a database of bovine herpesvirus type 1 (BHV1)-free farms to study the probability of, and risk factors for, introduction of BHV1, bovine viral diarrhoea virus (BVDV), Salmonella enterica subsp. enterica serotype Dublin (S. dublin), and Leptospira interrogans serovar hardjo (L. hardjo).Although most of the 95 SPF farms had a low risk on introduction of infectious diseases, one disease was introduced into 12 farms and two diseases were introduced into one farm. Three farms experienced an outbreak of BHV1, one farm an outbreak of L. hardjo, two farms BVDV, six farms S. dublin, and one farm both BHV1 and S. dublin. The total incidence rate was 0.09 (0.06-0.12) per herd-year at risk. The results suggest that the "non-outbreak" farms were significantly more closed than the "outbreak" farms. Direct animal contacts with other cattle should be avoided and professional visitors should be instructed to wear protective clothing before handling cattle.     

畜产品安全中存在的问题与分析

阐述了我国畜产品安全现状,强调了其形势的严峻性和紧迫性,分析了当前存在的主要问题,并提出了改进措施和建议。     

Cow-level evaluation of a kinetics ELISA with multiple cutoff values to detect fecal shedding of Mycobacterium avium subspecies paratuberculosis in New York State dairy cows

In control programs for Mycobacterium avium subsp. paratuberculosis (Map), the infection status of the cows in a herd is often obtained by testing (a sample of) the herd with an ELISA that may lack some sensitivity and specificity but that is fast and inexpensive. In New York State (NYS), an unabsorbed kinetics ELISA (KELA) has been used extensively for Map control. The objective of this study was to determine the relative sensitivity and specificity of the KELA for detection of fecal shedding of Map for the NYS dairy cow population, taking into account possible confounders such as different antigen batches and Map prevalence in a herd.

The data for the study consisted of all serum samples from NYS dairy cows with concurrent fecal culture results submitted to the NY Animal Health Diagnostic Laboratory (NYAHDL) between 1991 and 1996 (n = 10,562). The data represented cows with different levels of fecal shedding from herds with different within-herd Map prevalence, including herds that were whole herd fecal culture negative on repeated testing.

The cutoff values were based on the predictive value for fecal shedding obtained with a multiple logistic regression model that included variables for the three antigen batches and the Map prevalence in the herd. The KELA could not distinguish between non-shedders and low shedders (≤30 total colony forming units (TCFU)) and thus the predictive value of the KELA to detect moderate to heavy fecal shedders (>30 TCFU) was modeled. The three cutoff values of 65, 135 and 170 were based on low (<0.2), moderate (<0.80) and high (>0.95) probabilities for moderate to heavy fecal shedding. The sensitivity and specificity values relative to culture were 67% and 95.2%, 31% and 99.7%, and 11% and 99.9% for the three cutoff values, respectively. Cutoff values for the KELA decreased for herds with increasing within-herd Map prevalence. For the best positive predictive value of a KELA for moderate to heavy fecal shedding, the cutoff values should be determined based on the apparent within-herd prevalence in a herd.     

The use of Markov chain Monte Carlo for analysis of correlated binary data: patterns of somatic cells in milk and the risk of clinical mastitis in dairy cows

Two analytical approaches were used to investigate the relationship between somatic cell concentrations in monthly quarter milk samples and subsequent, naturally occurring clinical mastitis in three dairy herds. Firstly, cows with clinical mastitis were selected and a conventional matched analysis was used to compare affected and unaffected quarters of the same cow. The second analysis included all cows, and in order to overcome potential bias associated with the correlation structure, a hierarchical Bayesian generalised linear mixed model was specified. A Markov chain Monte Carlo (MCMC) approach, that is Gibbs sampling, was used to estimate parameters.

The results of both the matched analysis and the hierarchical modelling suggested that quarters with a somatic cell count (SCC) in the range 41,000–100,000 cells/ml had a lower risk of clinical mastitis during the next month than quarters <41,000 cell/ml. Quarters with an SCC >200,000 cells/ml were at the greatest risk of clinical mastitis in the next month. There was a reduced risk of clinical mastitis between 1 and 2 months later in quarters with an SCC of 81,000–150,000 cells/ml compared with quarters below this level. The hierarchical modelling analysis identified a further reduced risk of clinical mastitis between 2 and 3 months later in quarters with an SCC 61,000–150,000 cells/ml, compared to other quarters.

We conclude that low concentrations of somatic cells in milk are associated with increased risk of clinical mastitis, and that high concentrations are indicative of pre-existing immunological mobilisation against infection. The variation in risk between quarters of affected cows suggests that local quarter immunological events, rather than solely whole cow factors, have an important influence on the risk of clinical mastitis. MCMC proved a useful tool for estimating parameters in a hierarchical Bernoulli model. Model construction and an approach to assessing goodness of model fit are described.     

Effect of pathogen-specific clinical mastitis on herd life in two New York State dairy herds

The objective of this study was to estimate the effects of clinical mastitis (CM) (both with and without specific pathogen identification) occurring in different stages of lactation on length of herd life in two New York State dairy farms. The 2,697 cows in the study were followed for one lactation (the first-occurring one on or after 1 October 1999), until it ended because of a new lactation, culling, or end of study (31 March 2001 in one farm; 31 July 2001 in the other). A Cox proportional hazards model with time-dependent covariates, in SAS((R)), was used to measure, within a lactation, the effect of the first occurrence of CM (without specific pathogen identification) occurring 1--7, 8--66, 67--100, 101--225, or >or=226 days in milk (DIM), on how long cows remained in the herd. For the first occurrence of CM due to Streptococcus spp., Staphylococcus aureus, Staphylococcus spp., Escherichia coli, Klebsiella spp., and 'no pathogen isolated', the intervals were before and after the median DIM of first occurrence of each pathogen. There were too few cases due to Arcanobacterium pyogenes, and 'other pathogens grouped together' to split into intervals, so they were modeled as binary variables, i.e. as they occurred. CM was modeled using time-dependent covariates, to account for its differing effects throughout lactation on culling. Other variables controlled for were herd, parity, calving season, and other significant diseases. In the dataset, the lactational incidence risk of the first occurrence of CM was 18.2%; 20.0% of the cows did not survive the lactation that was studied. The overall annual culling percentage for both herds during the study period (including all cows, whether eligible for the study or not) was 35.6%. For cows with CM without pathogen identification, their highest hazard ratio (HR) of culling occurred from 67 to 100 DIM. All of the pathogens modeled markedly reduced herd life. On average over the entire lactation, cows with Staphylococcus spp. CM had the highest HRs for culling, although there were no significant differences among pathogens (at p=0.0018 (reflecting 28 pairwise comparisons)). For early-occurring (before median DIM of first occurrence) S. aureus CM, the daily rate of change of the HR of culling increased over time. The HRs for culling were particularly high for late-occurring (after median DIM of first occurrence) E. coli and Klebsiella spp. CM early in the interval, but the daily rate of change of the hazard of culling for these two pathogens decreased sharply over time. Treating CM as time-dependent therefore allowed us to measure in greater detail, its varying effects (of when it occurred) on herd life.     

Antimicrobial susceptibility of coagulase-negative staphylococci isolated from bovine milk samples

The aim of this study was to examine whether antimicrobial resistance profiles of coagulase-negative Staphylococcus (CNS) species isolated from milk of dairy cows differed between bacterial species, and to compare results obtained by phenotypic and genotypic profiling of resistance to penicillin, oxacillin and macrolide-lincosamide (ML) antibiotics. Of 170 CNS isolates, 83 (48.8%) were phenotypically susceptible to all antimicrobial agents tested in minimum inhibitory concentration (MIC) assays, 40.6% expressed resistance to a single compound or a single class of compounds, and 10.6% to multiple drug classes. Nine percent, 68%, 19%, 4% and 1% of isolates were negative for all resistance genes tested by PCR or positive for one, two, three or four resistance genes, respectively. Phenotypic resistance and detection of resistance genes other than blaZ were relatively rare in Staphylococcus chromogenes, which was the most common CNS species (36% of 170 genotypically identified isolates). In Staphylococcus epidermidis, which was the second most common CNS species (14% of isolates), phenotypic penicillin resistance was significantly more common than in other CNS species. Almost half of the S. epidermidis isolates carried multiple resistance genes and 30% carried the methicillin resistance gene mecA. Survival analysis using MIC values showed significant associations between phenotypic and genotypic resistance profiles. We conclude that CNS species from bovine milk differ significantly in phenotypic and genotypic antimicrobial resistance profiles, which has implications for treatment and management decisions.     

Antimicrobial Resistance Impacts Clinical Outcome of Granulomatous Colitis in Boxer Dogs

Background: Escherichia coli have recently been identified within the colonic mucosa of Boxer dogs with granulomatous colitis (GC). Eradication of invasive E. coli is associated with clinical and histological remission. Objectives: To determine antimicrobial susceptibility profiles of E. coli strains from GC and healthy dogs, and the association of antimicrobial resistance with clinical outcome. Animals: Fourteen Boxer dogs with GC and 17 healthy pet dogs. Methods: Prospective study: E. coli was cultured from GC biopsies and rectal mucosal swabs of healthy dogs. Individual strains were selected by phylogroup and overall genotype, determined by triplex‐ and random amplified polymorphic DNA‐polymerase chain reaction respectively. Antimicrobial susceptibility was determined by broth microdilution minimal inhibitory concentration. Results: Culture yielded 23 E. coli strains from GC (1–3/dog, median 2) and 34 strains from healthy (1–3/dog, median 2). E. coli phylogroups were similar (P= .18) in GC (5A, 7B1, 5B2, 6D) and healthy (2A, 10B1, 15B2, 7D). Resistance to ampicillin, amoxicillin‐clavulanate, cefoxitin, tetracycline, trimethoprim‐sulfa (TMS), ciprofloxacin, and chloramphenicol was greater (P < .05) in GC (21–64%) than healthy (0–24%). Enrofloxacin resistant E. coli were isolated from 6/14 GC versus 0/17 healthy (P= .004). Of the enrofloxacin resistant cases, 4/6 were also resistant to macrophage‐penetrating antimicrobials such as chloramphenicol, rifampicin, and TMS. Enrofloxacin treatment before definitive diagnosis was associated with antimicrobial resistance (P < .01) and poor clinical outcome (P < .01). Conclusions and Clinical Importance: Antimicrobial resistance is common among GC‐associated E. coli and impacts clinical response. Antimicrobial therapy should be guided by mucosal culture and antimicrobial susceptibility testing rather than empirical wisdom.