Histophilus somni infection in cows with fertility problems and vaginitis

Histophilus somni associated with vaginitis in cattle; fasciolosis still prevalent in cattle; clostridial metritis in a large goat herd; detachment of the ischial tuberosity in pigs; fowl cholera in commercial and backyard flocks. These are among matters discussed in the Veterinary Laboratories Agency's (VLA's) disease surveillance report for September.     

Single nucleotide polymorphisms in the bovine Histophilus somni genome; a comparison of new and old isolates

Histophilus somni, a causative agent of the bovine respiratory disease complex, can also cause a variety of systemic disorders, including bronchopneumonia, myocarditis, pericarditis, arthritis, pleuritis, and infectious thrombotic meningoencephalitis. The purpose of this study was to determine if currently circulating strains differ from those of the 1980s by identifying genomic changes. Single nucleotide polymorphisms (SNPs) and insertion and deletion (INDEL) sites were examined by whole-genome sequencing in 12 samples, 6 old and 6 new. The 31 028 SNP/INDELs recorded were compared against the reference genome sequence of the pathogenic H. somni strain 2336. The distribution of about 75% of these SNPs within a specified gene differed between old and new isolates and did not follow any particular pattern. The other 25% clustered into 2 groups containing the same SNPs in various genes: group I included 5 old isolates and 1 new isolate; group II included 5 new isolates and 1 old isolate. For putative virulence genes there were more SNPs in group I compared with strain 2336, itself an older isolate, than in group II. Although only 25% of all the SNPs formed 2 clusters, the results suggest some genetic difference in various genes between old and new strains.     

Coxiella burnetii Shedding During the Peripartum Period and Subsequent Fertility in Dairy Cattle

The objective of this study was to assess the effects of Coxiella burnetii shedding or seropositivity on post‐partum recovery and subsequent fertility in high‐producing dairy cows. Given the difficulty in diagnosing C. burnetii infection at the farm level, an exhaustive series of tests in 43 pregnant animals that delivered at least one live calf were conducted, including blood serology and PCR of milk or colostrum, cotyledons (only at parturition), faeces, vaginal fluid against C. burnetii on gestation Day 171–177, at parturition and on Days 1–7, 8–14, 15–21, 22–28, 29–35 and 90–97 post‐partum. During scheduled herd visits, ultrasonography (US) of the genital tract and examination of vaginal fluid were performed on Days 15–21 (V1), 22–28 (V2), 29–35 (V3) and 51–57 (V4) post‐partum by the same veterinarian. Logistic regression analysis revealed that the likelihood of suffering endometritis (the presence of echogenic intrauterine fluid (IUF), cervical diameter of ≥4 cm or endometrial thickness ≥0.75 cm) was lower in C. burnetii‐seropositive animals (OR = 0.10), compared with C. burnetii‐seronegative animals. According to Kaplan–Meier survival analysis, C. burnetii‐seronegative and non‐shedding cows showed a delayed return to luteal activity and conception was delayed in non‐shedding animals, compared with the remaining animals. Overall, the results of our study provide useful insight into the effects of C. burnetii infection on post‐partum recovery and subsequent fertility. In particular, animals not infected with Coxiella seem to be susceptible to infection and not protected against the bacterium in dairy herds. The elevated costs of determining an infection at the farm level, make monitoring of cows virtually impossible from a clinical point of view.     

关于奶牛在性周期不同阶段超排方法的研究

牛的胚胎移植技术在畜牧业发达国家已从实验阶段进入生产应用阶段.我国经近十币年的研究,也取得了很大进展.     

The Incidence and Significance of Bovine Herpesvirus (infectious bovine rhinotracheitis) Antibodies in the Sera of Aborting Cattle

The results of bovine herpesvirus (IBR-IPV) neutralization tests conducted on the sera of 463 animals with a prior history of undiagnosed abortion and 331 control animals with no history of abortion are reported. One hundred and thirty-one (28.3 per cent) of the aborting and 105 (31.7 per cent) of the control animals, were found to harbour antibody to this virus. No significant seasonal or gestational incidence could be determined but some variation in the annual incidence was apparent. On the basis of these results it appears unlikely that this virus is responsible for a significant proportion of the undiagnosed cases of bovine abortion in Quebec and Ontario, where the animals tested were located. The problems involved in substantiating a diagnosis of abortion due to bovine herpesvirus are briefly discussed.     

Potential Risk Factors Associated with Bovine Spongiform Encephalopathy in Cattle from Schleswig‐Holstein,Germany

Since the first detection of bovine spongiform encephalopathy (BSE) in homebred cattle in Germany on 26 November 2000, 382 cases have been confirmed until 30 September 2005. Thirty‐two of these cases were reported from the Federal State of Schleswig‐Holstein (SH). There are hypotheses on the routes of infection for German cattle, but only few efforts have been made to assess potential risk factors by epidemiological studies. The purpose of this study was to identify potential risk factors at the farm level for the occurrence of BSE in cattle in SH. By applying the method of indirect standardization, the prevalence of various structural and management parameters of BSE‐affected farms in SH was compared with the prevalence of these parameters in a standard population of cattle farms from SH. The data describing the standard population were obtained by a cross‐sectional study performed in SH in 2003. Data of the BSE case population were available from the central German BSE case database. A possible association of the occurrence of BSE with the feeding of milk replacers to calves was observed. There was a clear indication that the occurrence of BSE was associated with the presence of pigs and/or poultry on the farm. This finding suggests that cross‐contamination of feed or cross‐exposition may have occurred in SH. The results obtained using the indirect standardization analysis were validated by multiple logistic regression. This study indicates that the feedborne path has been the principal route of transmission for the BSE agent in SH.     

Therapeutic Approaches to Pregnancy Loss of Non‐infectious Cause During the Late Embryonic/Early Foetal Period in Dairy Cattle. A Review

The problem of pregnancy loss during the late embryonic/early foetal period due to non‐infectious causes in dairy cattle is on the rise, especially in high milk producing cows. This review discusses recent therapeutic approaches designed to reduce the incidence of early foetal loss. These strategies are based on the fact that low plasma concentrations of progesterone are commonly related to foetal loss. Progesterone supplementation during the late embryonic/early foetal period has proven useful in curtailing losses, mainly in single pregnancies, whereas a more positive effect of treatment with GnRH than progesterone has been found in twin pregnancies. Therapeutics is not necessary in cows with an additional corpus luteum. In practical terms, suggestions include recording the number of embryos and corpora lutea on first pregnancy diagnosis and checking for normal pregnancy progression on day 60 post‐insemination.     

Investigations on the Prevalence of Bovine Tuberculosis and Brucellosis in Dairy Cattle in Dar es Salaam Region and in Zebu Cattle in Lugoba Area,Tanzania

A study between August 1995 and December 1997 included 343 dairy cattle on 20 farms in the Dar es Salaam region and 2289 zebu cattle on 39 bomas in the Lugoba area (coast region). The aim was to establish the prevalence of bovine tuberculosis (Mycobacterium bovis) and bovine brucellosis (Brucella abortus). In the single intradermal tuberculin test (SIT), 0.9% (3/343) of the animals in Dar es Salaam tested positive and 1.2% (4/343) were doubtful. Positive reactors were found in 10% (2/20) of the farms. In the Lugoba area, 0.6% (14/2206) were positive and 6.8% (149/2206) doubtful, positive cases being found in 21% (8/39) of all bomas. In the slow agglutination test (SAT) for B. abortus, 14.1% (48/341) of the serum samples reacted positively in Dar es Salaam and 2.3% (8/341) were doubtful. Positive SAT reactors were identified on 25% (5/20) of the dairy cattle farms. In the Lugoba area, 12.3% (273/2221) proved to be positive SAT reactors and doubtful reactions were observed in 2.9% (64/2221). SAT-positive animals were detected on 87% (34/39) of all bomas. The prevalence in single herds in Dar es Salaam varied from 4.3% to 5.3% for the SIT and from 2.2% to 50% for the SAT. The prevalence in single herds in Lugoba area was between 1.1% and 2.9% for SIT and from 1.4% up to 62.1% for SAT. The two cattle populations differed significantly (p<0.001) in the prevalence of both bovine tuberculosis and bovine brucellosis. Two cows that were positive reactors were slaughtered and subjected to post-mortem examination, and organ samples were bacteriologically cultured. The occurrence of Mycobacterium tuberculosis was confirmed by polymerase chain reaction (PCR) in both cows.     

Epidemiological and Bacteriological Aspects of Mastitis Associated with Yellow‐Jacket Wasps (Vespula germanica) in a Dairy Cattle Herd

The German wasp, Vespula germanica has been observed to injure teats of dairy cows, causing lesions that are associated with clinical and subclinical mastitis. The presence of skin lesions on the teats, caused by the wasps, was recorded in a dairy cattle herd located in the Samaria foothills during July–October 1999. Wasp‐inflicted injuries were found in 43.6% (58 of 133) of the adult cows and 1.4% (one of 71) of the first‐calving cows. They were located in 42.4% of cows (n = 25) on all quarters, 18 cows (30.5%) had lesions on the front quarters and 27.1% (n = 16) of cows on the hind quarters only. Clinical and subclinical mastitis were diagnosed in 61% (36 of 59) and 28.8% (17 of 59), respectively, of the injured adult and first‐calving cows. The most common bacterial isolates from the mastitic cows were Staphylococcus aureus 45.1% (n = 14), Streptococcus dysgalactiae 16.1% (n = 5), Streptococcus spp. 19.4% (n = 7) and others 13.9% (n = 5). The loss of milk production was estimated at 300 kg milk for each cow injured by wasps and exhibiting clinical mastitis. An increase in the bulk‐milk somatic cell count, from 186 × 10³ at 1 month prior to the outbreak to a peak of 1200 × 10³ in the post‐outbreak month, was noted. The culling rate reached 13.6% (eight of 59) of the affected cows. In summary, the considerable economic losses caused by the wasp infestation resulted from decreased milk production and a decline in milk quality, culling of affected cows, and increased demand for use of drugs and veterinary care.     

Bovine Herpesvirus 5 (BoHV-5) in Bull Semen: Amplification and Sequence Analysis of the US4 Gene

Bovine herpesvirus type 5 (BoHV-5), which is potentially neuropathogenic, was detected in clinical samples of bovine semen, both directly and after isolation in cell culture, using a nested PCR system for amplifying the US4 gene. Nucleotide sequences generated from the amplicons were analysed and deposited at GenBank (NCBI, Bethesda, MD, USA) under the accession numbers AF298174 and AF330157. Alignment of these sequences and previously deposited sequences of BoHV-1 and BoHV-5 showed 82% and 98% similarity, respectively. The bulls, which were maintained at an artificial insemination centre, had presented no clinical signs, indicating that bovine semen should be screened for BoHV-5 to prevent transmission of the virus.     

Effects of Co‐culture of Cumulus Oocyte Complexes with Denuded Oocytes During In Vitro Maturation on the Developmental Competence of Cloned Bovine Embryos

This study evaluated the effects of co‐culture of immature cumulus oocyte complexes (COCs) with denuded immature oocytes (DO) during in vitro maturation on the developmental competence and quality of cloned bovine embryos. We demonstrated that developmental competence, judged by the blastocyst formation rate, was significantly higher in the co‐cultured somatic cell nuclear transfer (SCNT+DO, 37.1 ± 1.1%) group than that in the non‐co‐cultured somatic cell nuclear transfer (SCNT‐DO, 25.1 ± 0.9%) group and was very similar to that in the control IVF (IVF, 38.8 ± 2.8%) group. Moreover, the total cell number per blastocyst in the SCNT+DO group (101.7 ± 6.2) was higher than that in the SCNT‐DO group (81.7 ± 4.3), while still less than that in the IVF group (133.3 ± 6.0). Furthermore, our data showed that mRNA levels of the methylation‐related genes DNMT1 and DNMT3a in the SCNT+DO group were similar to that in the IVF group, while they were significantly higher in the SCNT‐DO group. Similarly, while the mRNA levels of the deacetylation‐related genes HDAC2 and HDAC3 were significantly higher in the SCNT‐DO group, they were comparable between the IVF and SCNT+DO groups. However, the mRNA levels of HDAC1 and DNMT3B were significantly higher in the SCNT+DO group than in the other groups. In conclusion, the present study demonstrated that co‐culture of COCs with DO improves the in vitro developmental competence and quality of cloned embryos, as evidenced by increased total cell number.     

妊娠后期奶牛酮病对静脉血液气体和酸碱值影响的研究

本试验分析了17头妊娠后期患酮病奶牛和19头健康奶牛的十二项颈静脉血液气体和酸碱值指标。结果,酮病奶牛静脉血液气体和酸碱值中,酸碱度、氧分压、缓冲碱、氧饱和度、氧含量五项指标均数与健康奶牛相比无明显变化(p>0.05);而二氧化碳分压(PCO_2)为38.2±12.5mmHg,剩余碱(BE)-1.9±4.3mmol/l,细胞外液剩余碱(BEecf)为-2.7±5.7mmol/l,实际碳酸氢盐(HCO_3~-)为21.5±6.3mmol/l,二氧化碳总量(CO_2tot)为22.6±6.6mmol/l,标准碳酸氢盐(HCO_3~-St)为21.6±4.0mmol/l,标准酸碱度(pHst)为7.346±0.093,以上七项指标均数与健康奶牛相比明显降低(p<0.05),说明妊娠后期奶牛酮病对它们的影响较大,认为可做为综合诊断奶牛酮病和掌握妊娠后期奶牛代谢状态的依据。     

Plasma Concentrations of Pregnancy‐Associated Glycoproteins Measured Using Anti‐Bovine PAG‐2 Antibodies on Day 120 of Gestation Predict Abortion in Dairy Cows Naturally Infected with Neospora caninum

The present study sought to determine: (i) the effects of Neospora caninum infection and twin pregnancy on plasma pregnancy‐associated glycoprotein‐2 (PAG‐2) concentrations throughout pregnancy and (ii) whether plasma PAG‐2 concentrations could predict abortion in N. caninum‐infected cows. The study was performed on a commercial Holstein‐Friesian dairy herd in northeastern Spain and the final data included those recorded in 53 non‐aborting and 19 aborting animals. Blood samples were collected immediately before pregnancy diagnosis (on Days 40, 90, 120, 150, 180 and 210 post‐insemination) in non‐aborting cows or until the time of abortion detection in aborting cows. General lineal models (GLM) repeated measures anova revealed the different behaviour of PAG‐1 and PAG‐2, and significant effects of Neospora seropositivity, cool season and twin pregnancy on plasma PAG‐2 concentrations throughout gestation (between‐subject effects). In addition, based on the odds ratios, the likelihood of abortion increased in Neospora‐seropositive cows (by a factor of 7.0) compared to seronegative animals and decreased in cows with a high plasma PAG‐2 concentration (>4.5 ng/ml) on Day 120 of pregnancy (by a factor of 0.24), compared to the remaining cows. In conclusion, there is a relationship between plasma PAG‐2 concentrations and the risk of abortion in Neospora‐infected dairy cows. Thus, plasma PAG concentrations measured using anti‐boPAG‐2 antiserum on Day 120 of gestation could serve as an indicator of the abortion risk in N. caninum infected animals; values <4.5 ng/ml indicating a high risk of abortion in chronically infected animals.     

Pregnancy Loss in Dairy Cattle: Relationship of Ultrasound,Blood Pregnancy‐Specific Protein B,Progesterone and Production Variables

Objectives were to determine associations between percentage pregnancy loss (PPL) in dairy cattle and: (i) pregnancy diagnosis by ultrasonography; (ii) pregnancy diagnosis by serum pregnancy‐specific protein B (PSPB) concentrations, with or without serum progesterone concentrations; and (iii) production and environmental factors. This study included 149 822 pregnancy diagnoses conducted over 13 years in Holstein‐Friesian cows in Hungarian dairy herds. The following were determined: PPL in cows diagnosed pregnant by transrectal ultrasonography 29–42 days after artificial insemination (AI; n = 11 457); PPL in cows diagnosed pregnant by serum PSPB 29–35 days after AI (n = 138 365); and PPL and its association with serum progesterone concentrations, PSPB and production/environmental variables. The definition of PPL was percentage of cows initially diagnosed pregnant based on ultrasonography or PSPB, but not pregnant when examined by transrectal palpation 60 –70 days after AI. The PPL was lower (p < 0.001) in cows following ultrasonographic vs PSPB diagnosis of pregnancy at 29–35 days (8.1 vs 19.3%, respectively), but was higher in cows following ultrasonographic pregnancy diagnosis on 29–35 vs 36–42 days (8.1 vs 7.1%, respectively, P < 0.05). Furthermore, 72.9% of pregnancies with ultrasound‐detected morphological abnormalities resulted in pregnancy loss. As a subset of PSPB data, a fully quantitative PSPB assay was used for 20 430 samples; PPL in cows with a high PSPB concentration (>1.1 ng/ml) was lowest (15.0%), whereas cows with low concentrations of both PSPB and progesterone (0.6–1.1 and <2 ng/ml, respectively) had the highest PPL (76.3%; p < 0.0001). Furthermore, PPL was higher in cows with advanced parity and with high milk production, when ambient temperatures were high, although body condition score (BCS) had no effect on PPL. Finally, there were no significant associations between serum PSPB and environmental temperatures or number of post‐partum uterine treatments.     

Kisspeptin,c‐Fos and CRFR Type 2 Co‐expression in the Hypothalamus after Insulin‐Induced Hypoglycaemia

Normal reproductive function is dependent upon availability of glucose and insulin‐induced hypoglycaemia is a metabolic stressor known to disrupt the ovine oestrous cycle. We have recently shown that IIH has the ability to delay the LH surge of intact ewes. In the present study, we examined brain tissue to determine: (i) which hypothalamic regions are activated with respect to IIH and (ii) the effect of IIH on kisspeptin cell activation and CRFR type 2 immunoreactivity, all of which may be involved in disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and at 28 h after progesterone withdrawal (PW), animals received saline (n = 6) or insulin (4 IU/kg; n = 5) and were subsequently killed at 31 h after PW (i.e., 3 h after insulin administration). Peripheral hormone concentrations were evaluated, and hypothalamic sections were immunostained for either kisspeptin and c‐Fos (a marker of neuronal activation) or CRFR type 2. Within 3 h of treatment, cortisol concentrations had increased whereas plasma oestradiol concentrations decreased in peripheral plasma (p < 0.05 for both). In the arcuate nucleus (ARC), insulin‐treated ewes had an increased expression of c‐Fos. Furthermore, the percentage of kisspeptin cells co‐expressing c‐Fos increased in the ARC (from 11 to 51%; p < 0.05), but there was no change in the medial pre‐optic area (mPOA; 14 vs 19%). CRFR type 2 expression in the lower part of the ARC and the median eminence was not altered by insulin treatment. Thus, disruption of the LH surge after IIH in the follicular phase is not associated with decreased kisspeptin cell activation or an increase in CRFR type 2 in the ARC but may involve other cell types located in the ARC nucleus which are activated in response to IIH.     

Evaluation and Comparison of 2 On‐Farm Tests for Estimating Somatic Cell Count in Quarter Milk Samples from Lactating Dairy Cattle

Background

The somatic cell count (SCC) is commonly used to monitor udder health and diagnose subclinical intramammary infection (IMI) in dairy cattle.

Hypothesis

The Somaticell test (ST) 2 and California mastitis test (CMT) are clinically useful cow‐side tests for diagnosing subclinical IMI.

Animals

One hundred and eleven dairy cows at dry‐off and 92 cows within 4–7 days postcalving.

Methods

Quarter foremilk samples were obtained and analyzed with a DeLaval cell counter (DCC, reference method), 1 ST, and CMT. The ST was run in a simulated cow‐side manner using milk at 37°C instead of 0–8°C as recommended by the manufacturer. Test performance for diagnosing IMI (DCC SCC >200,000 cells/mL) was evaluated by calculating the area under the receiver operating characteristic curve (AUC) and the kappa coefficient (κ) at the optimal cut‐point for each test. The effect of milk/reagent temperature also was evaluated.

Results

Compared to the reference method, the ST run in a simulated cow‐side manner had an AUC = 0.68 and κ = 0.24 at dry‐off, and AUC = 0.74 and κ = 0.40 in fresh cows. The CMT performed much better than the ST in diagnosing subclinical IMI with AUC = 0.88 and κ = 0.77 at dry‐off, and AUC = 0.87 and κ = 0.76 in fresh cows. The measured ST value decreased with increasing temperature of the milk/reagent mixture.

Conclusions/Clinical Importance

The ST is optimized for use on milk at 0–8°C and is therefore designed for on‐farm use on refrigerated milk samples. The ST is not suited for use as a cow‐side screening test for IMI because the milk temperature exceeds the recommended range for the test.     

Effect of β‐Carotene Supply During Close‐up Dry Period on the Onset of First Postpartum Luteal Activity in Dairy Cows

The aim of this study was to examine the effect of β‐carotene supply during the close‐up dry period on the onset of first postpartum luteal activity in dairy cows. Twelve cows were supplied with 2000 mg of β‐carotene (20 g Rovimix^®β‐Carotene containing 10%β‐carotene; DSM Nutrition Japan K.K., Tokyo, Japan) by oral administration daily from day 21 before expected calving date to parturition. Fourteen cows (control) did not receive β‐carotene supplementation. Blood samples were obtained on days 21, 14 and 7 before expected calving date and on days 1, 7, 14, 21 postpartum. When the plasma progesterone concentration exceeded 1 ng/ml by day 21 postpartum, luteal activity was assumed to have been initiated. The result showed that serum β‐carotene concentrations in the β‐carotene cows were higher than in the control cows during the experimental period (p < 0.01). The number of cows with the onset of luteal activity by day 21 postpartum was 9/12 in the β‐carotene cows and 4/14 in the control cows (p < 0.05). Retinol, certain metabolic parameters and metabolic hormones concentrations did not differ between β‐carotene and control cows. In addition, serum retinol concentration in β‐carotene cows without luteal activity was lower than in β‐carotene cows with luteal activity (p < 0.05), and serum gamma‐glutamyl transpeptidase concentration in β‐carotene cows with luteal activity (p < 0.05) and control cows without luteal activity (p < 0.01) was higher than in control cows with luteal activity. In conclusion, β‐carotene supply during the close‐up dry period may support the onset of luteal activity during early lactation in dairy cows.     

Fertility in Single‐ovulating and Superovulated Dairy Heifers after Insemination with Low Dose Sex‐sorted Sperm

The present study was performed to test fertility in single‐ovulating and superovulated dairy heifers after insemination with low dose sex‐sorted sperm under field conditions. Some parameters, including the dosage, deposition site and timing, were assessed with the pregnancy rates after artificial insemination (AI). Moreover, the use of oestrus synchronization in combination with sorted sperm was evaluated. Besides that, we also improved the embryo production efficiency in superovulated dairy heifers by optimizing the timing of inseminations and repartitioning the sexed sperm dosage among multiple inseminations. The conception rate (52.8%) in heifers after low dose (2 × 10⁶) insemination with sorted sperm deep into the uterine horn did not differ (p > 0.05) from that (59.6%) of conventional AI (1 × 10⁷ non‐sorted sperm) and that of deep insemination with low dose non‐sorted sperm (57.7%). There was also no difference (p > 0.05) between conception rates after single (51.7%) and double (53.8%) deep insemination with sorted semen. Heifers inseminated with sorted sperm at synchronous oestrus had a lower pregnancy rate (48.1%) than heifers at spontaneous oestrus (53.6%), but this did not reach statistical difference (p > 0.05). The average number of transferable embryos collected in vivo from heifers inseminated with sorted sperm (4.81 ± 2.04) did not differ (p > 0.05) from that obtained from heifers after insemination with non‐sorted sperm (5.36 ± 2.74). Thus, we concluded that the pregnancy rate after deep intra‐uterine insemination with low dose sorted sperm was similar to that of non‐sorted sperm, which was either also deposited at a low dose deep intra‐uterine or into the uterine body. Sychronization of oestrus can be beneficial in combination with sorted sperm to optimize the organization and management of dairy herds. The results from superovulated heifers demonstrated that our insemination regime can be used to obtain a comparable embryo production efficiency with sorted sperm than with non‐sorted sperm.