Real‐time quantitative PCR‐based detection of Coxiella burnetii in unpasteurized cow's milk sold for human consumption

Coxiella burnetii is a zoonotic pathogen with a worldwide distribution that is responsible for Q fever in humans. It is a highly infectious bacterium that can be transmitted from cattle to humans through the consumption of unpasteurized milk. We report the molecular identification of C. burnetii in raw cow's milk being sold directly for human consumption in Brazil without official inspection or pasteurization. One hundred and twelve samples of raw milk were analysed by real‐time quantitative PCR (qPCR), and C. burnetii was detected in 3.57% (4/112) of the samples at a concentration ranging from 125 to 404 bacteria per millilitre. The identification of this zoonotic pathogen in raw milk sold directly for human consumption is a public health concern since C. burnetii can be transmitted through the oral route. This result indicates that health education and other preventive measures should be officially implemented in Brazil to prevent the spread of infection. To our knowledge, this is the first qPCR‐based detection of C. burnetii in raw milk samples from cows sold in Brazil that do not undergo official inspection or pasteurization.     

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.     

A Systematic Review and Meta‐Analysis of Phase I Inactivated Vaccines to Reduce Shedding of Coxiella burnetii From Sheep and Goats From Routes of Public Health Importance

Q fever in humans and coxiellosis in livestock are both caused by Coxiella burnetii. The public health importance of vaccination against C. burnetii shedding from sheep and goats was evaluated using systematic review and meta‐analysis to provide evidence for policy direction to prevent potential zoonotic spread. Publications reporting shedding of C. burnetii in vaginal and uterine secretions, milk, placenta and faeces were included. A single observational (one goat) and seven experimental (four goat and three sheep) vaccine studies were included in the review. No relevant publications on other interventions were identified. Random effects meta‐analyses were performed for the risk of shedding in individuals in the control and vaccinated groups and for the mean difference in the level of bacterial shedding in sheep and goats stratified by age and previous exposure status. Limited data were available for further analytic evaluation. From the pooled analysis, an inactivated phase I vaccine significantly reduced the risk of shedding from uterine (RR = 0.10; 95%CI 0.05–0.20) secretions in previously sensitized goats. Individual studies reported significant risk reduction in milk (RR = 0.03; 95%CI 0.01–0.26), vaginal secretions (RR = 0.40; 95%CI 0.22–0.75) and faeces (RR = 0.79; 95%CI 0.63–0.97) from naïve goats. The pooled mean levels of bacteria shed from placental [mean difference (MD = ?5.24 Log₁₀; 95%CI ?6.75 to ?3.7)] and vaginal (MD = ?1.78 Log₁₀; 95%CI ?2.19 to ?1.38) routes were significantly decreased in vaccinated naïve goats compared with controls. Shedding through all other routes from vaccinated goats was not significantly different than shedding from control goats. No effect of vaccination was found on the risk of shedding or the mean level of shedding in vaccinated sheep compared with control sheep. Our conclusions are based on a limited amount of data with variable risk of systematic error.     

Lack of effects of an equine chorionic gonadotropin (eCG) administration between days 9 and 15 postpartum on reproductive performance in a Holstein dairy herd

Recently, it has been demonstrated that administration of equine chorionic gonadotropin (eCG) in the postpartum period in dairy cows can enhance follicle growth, reduce the interval from calving to first ovulation and increase plasma estradiol concentrations, and, thus, could enhance reproductive performance in a dairy herd when administered on day 6 postpartum. The aim of this study was to investigate the effects of a single dose of eCG between days 9 and 15 postpartum on parameters of reproductive performance in dairy cows. German Holstein cows (n = 1937; primiparous cows: n = 748; pluriparous cows: n = 1189) in a commercial dairy farm were randomly assigned to three experimental groups. Animals within the group eCG received a single dose of 600 IU eCG intramuscularly (i.m.) between days 9 and 15 postpartum followed by an i.m. administration of 500 μg cloprostenol after 14 days. Those of treatment group PG received cloprostenol only between days 23 and 29 postpartum. Cows of the control group remained untreated. Starting on day 49 postpartum, cows were subjected to a Presynch‐Ovsynch protocol and inseminated artificially. The impact of application time (days postpartum) of eCG on the intervals calving to first service and calving to conception was statistically not significant. Outcomes of reproductive performance (i.e. first service conception rate, proportion of pregnant cows until 100 and 150 days in milk [DIM], number of inseminations until 150 DIM, calving to first service interval and calving to conception interval) did not differ significantly between treatment group eCG and group PG compared to control group. Regarding postpartum eCG administration, significant interactions between treatment and parity, season, milk yield, and early puerperal disorders, respectively, could not be shown. In conclusion, an eCG treatment of dairy cows between days 9 and 15 postpartum to increase reproductive performance cannot be recommended under the given circumstances.     

Effect of parity number on the dry matter intake of dairy cows during the first week after calving

We investigated the effect of parity number on the dry matter intake (DMI) of cows during the first week after calving. Eighty‐three cows were evaluated from 14 days before to 7 days after calving. DMI and milk yield were measured for 7 days after calving, and the calving score was measured. Blood samples were collected throughout the experiment. The average DMI during the first week after calving was reduced in the first‐lactation heifers and high‐parity number cows. A quadratic relationship between the parity number and the DMI was observed. The first‐lactation heifers had lower prepartum serum total protein (TP) concentration and milk yield, higher prepartum serum nonesterified fatty acid (NEFA) concentration and calving score than the multiparous cows. The recovery rate of serum calcium (Ca) after calving was slow in the cows in the parity 6. The DMI was positively affected by the serum Ca concentration after calving, milk yield, and prepartum serum TP concentration and was negatively affected by the calving score and prepartum serum NEFA concentration. We conclude that the DMI immediately after calving tends to be lower in first‐lactation heifers and high‐parity number cows, but factors that reduce the DMI differ according to parity number.     

Seroprevalence of Coxiella burnetii in Australian dogs

The role of dogs in the transmission of Coxiella burnetii to humans is uncertain, and extensive seroprevalence studies of dogs have not been previously conducted in Australia. This study determined C. burnetii exposure in four diverse canine subpopulations by adapting, verifying and comparing an indirect immunofluoresence assay (IFA) and an enzyme‐linked immunosorbent assay (ELISA) used to detect anti‐C. burnetii antibodies in humans. Canine serum samples (n = 1223) were tested with IFA from four subpopulations [breeding establishments; household pets; free‐roaming dogs in Aboriginal communities; shelter dogs]. The proportions of seropositive dogs were as follows: breeding (7/309, 2.3%), household pets (10/328, 3%), Aboriginal communities (21/321, 6.5%) and shelters (5/265, 1.9%). Dogs from Aboriginal communities were 2.8 times (CI 1.5–5.1; P < 0.001) more likely to be seropositive than dogs from other populations. The ELISA was used on 86 of 1223 sera tested with IFA, and a Cohen's Kappa coefficient of 0.60 (CI 0.43–0.78) indicated good agreement between the two assays. This study has established that Australian dogs within all four subpopulations have been exposed to C. burnetii and that a higher seroprevalence was observed amongst free‐roaming dogs associated with Aboriginal communities. As C. burnetii recrudesces during pregnancy and birth products contain the highest concentration of organism, individuals assisting at the time of parturition, those handling pups shortly after birth as well as those residing in the vicinity of whelping dogs are potentially at risk of developing Q fever. However, the identification of active antigen shed in excreta from seropositive dogs is required in order to accurately define and quantify the public health risk.     

Disease Risk Surface for Coxiella burnetii Seroprevalence in White‐Tailed Deer

Coxiella burnetii is considered a re‐emerging zoonosis in many countries. The bacterium is enzootic in livestock and wildlife in the United States, and environmental contamination is widespread. Despite the potential for exposure, the estimated prevalence of Q fever in humans and animals is not well elucidated, and reported human infections in the United States are relatively rare. Zoonotic transmission of the bacterium is usually associated with abortions in domestic ruminants, but other modes of transmission, such as contact with infected blood and/or milk during field dressing of infected wildlife, have not been thoroughly investigated. Studies of zoonotic pathogen transmission between animal reservoir hosts and humans are usually established in response to documented emergence or re‐emergence of a zoonosis in a particular locale, and, as such, the prevalence of infection in wildlife is largely unknown for many zoonotic pathogens, including C. burnetii. The objective of this study was to create a disease risk surface for C. burnetii seroprevalence in wild white‐tailed deer (Odocoileus virginianus) in New York State. Blood samples were collected from hunter‐harvested deer from across New York State in 2009 and 2010. The samples were processed and tested for the presence of anti‐C. burnetii antibodies via indirect microimmunofluorescence assays using phase II C. burnetii strain RSA439. Overall, 14.50% of the tested white‐tailed deer were C. burnetii phase II seropositive. The dual Kernel density estimation method was used to create a smoothed disease risk surface, which revealed variation in seroprevalence ranging from 0% to 32.0%. Areas of higher seroprevalence were detected in four discrete areas of Central New York and in one additional area in the southwest corner of the northern part of the state. This suggests certain locales where humans may be at increased risk for exposure to the bacterium secondary to contact with potentially infected deer.     

Effects of pre‐partum milking of dairy cows on calcium metabolism at start of milking and at calving

This experiment studied the effect of pre‐partal milk removal on calcium metabolism at start of milking and at calving. Nine cows of the Swedish Red breed were milked for 1–7 days pre‐partum. The average milk yield at the first milking was 4.8 l, and the average yield the last day prior to calving was 13.4 l. Five cows were used as control cows and were only milked post‐partum. Samples of plasma and urine were taken to determine the effect of pre‐partum milking and calving on levels of calcium, magnesium, parathyroid hormone and plasma C‐terminal crosslinked telopeptide of type 1‐collagen (CTx), used as a marker of bone resorption. Pre‐partum milking resulted in a decrease in plasma calcium that was evident 2 days after the first milking. Parathyroid hormone increased at the same time, and CTx started to increase from 24 h after the first milking. There were no effects on plasma magnesium or urinary output of calcium or magnesium. The first week after calving, there were no differences between pre‐partum milked cows and control cows in plasma or urine variables, or in milk yield. In conclusion, pre‐partum milking activated the calcium‐restoring mechanisms but did not improve calcium status at calving.     

Peripartal calcium homoeostasis of multiparous dairy cows fed rumen‐protected rice bran or a lowered dietary cation/anion balance diet before calving

Milk fever is one of the most important metabolic diseases in dairy cattle. Reducing the dietary cation/anion balance (DCAD) with anionic salts is a common prevention strategy. However, many small European farms cannot use total mixed rations (TMR) in the close‐up period. Including anionic salts in compound feeds can result in feed refusals and moderate inclusions to preserve feed palatability results in insufficient DCAD reduction. Rumen‐protected rice bran induces the adaptation of Ca metabolism in dairy cows by a reduction of Ca intake and by a reduction of the availability of dietary Ca. In the presence of a negative control, rumen‐protected rice bran (2.8 kg/day) was compared with a lowered DCAD diet (from 269 to 4 meq/kg DM) in their effect to prevent milk fever. In a randomized block design, 45 multiparous Holstein cows joined the trial sequentially from 21 days before the expected calving date and were observed until the 8th week of lactation. Feed and nutrient intakes were recorded, and Ca, P, Mg in serum and urine, urine pH, serum NEFA and milk production in early lactation were compared. Feeding rumen‐protected rice bran before calving improved the recovery of calcaemia after calving and had a positive effect on DMI after calving. The moderately low DCAD diet did not positively influence serum Ca at calving. Calcaemia recovered even later than in control, and cows showed reduced DMI post‐calving and higher NEFA levels in the first 36 h after calving. This moderate reduction of DCAD did not provide an intermediate prevention level indicating that DCAD needs to be reduced to the recommended levels to prevent milk fever. Rumen‐protected rice bran may be a suitable feed to reduce hypocalcaemia post‐partum and can be included in pre‐calving compound feeds representing a palatable alternative to anionic salts.     

Molecular detection of Coxiella burnetii in raw meat intended for pet consumption

The discovery of antibodies against Coxiella burnetii in cattery‐confined breeding cats indicating prior or current exposure (Shapiro et al., 2015) prompted an investigation into possible sources of infection. One hypothesis was that raw meat diets containing reservoir species may provide a source of C. burnetii transmission. The aim of this pilot study was to determine whether C. burnetii DNA was present in raw meat sold exclusively for companion animal consumption. The sample population consisted of raw meat packages (n = 58) of primarily kangaroo origin, with three to four aliquots (50–120 mg) randomly selected from each package. Genomic DNA was extracted from whole tissue in each of these aliquots using a modified protocol. Three quantitative PCR assays were used for the detection of C. burnetii targeting the IS1111 gene, the heat shock operon htpAB and the C. burnetii outer membrane protein‐coding gene, com1. Coxiella burnetii DNA was detected in 25/58 samples (43%) using the IS1111, htpAB and/or com1 PCR assays and confirmed by DNA sequencing. All samples amplifying a product in the com1 assay also amplified a product in the htpAB and IS1111 assays. A total of 17/58 (29%) packets were positive with all three genes, 4/58 (7%) were positive with two genes (IS1111 and htpAB) and 4/58 (7%) were positive with the IS1111 gene only. Coxiella burnetii DNA was five times more likely to be found in offal than skeletal muscle meat samples. All meat samples in which C. burnetii DNA was found were from kangaroo tissues, while samples labelled as non‐kangaroo meat (n = 4) were negative. Multi‐locus variable number of tandem repeat analysis (MLVA) identified three different genotypes of C. burnetii that have all been identified previously from Australian human clinical Q fever cases. Further investigations are required to determine the potential role of certain raw meats in the transmission of C. burnetii to cats and humans.     

Effects of Moringa oleifera silage on milk yield,nutrient digestibility and serum biochemical indexes of lactating dairy cows

This study investigated the effects of Moringa oleifera (MO) as a partial substitute of alfalfa hay on milk yield, nutrient apparent digestibility and serum biochemical indexes of dairy cows. MO was harvested at 120 days post‐seeding. Fresh MO was cut, mixed with chopped oat hay (425:575 on a DM basis), ensiled and stored for 60 days. Sixty healthy Holstein dairy cows were allocated to one of three groups: NM (no MO or control), LM (low MO; 25% alfalfa hay and 50% maize silage were replaced by MO silage) or HM (high MO; 50% alfalfa hay and 100% maize silage were replaced by MO silage). The feeding trial lasted 35 days. The LM and HM diets did not affect dry matter (DM) intake, milk yield or milk composition (lactose, milk fat, milk protein and somatic cell count). The apparent digestibility of DM and NDF was lower for HM group than NM group. Additionally, there were no significant differences in serum biochemical indexes between the LM and NM groups. The HM group had lower serum concentrations of total cholesterol, high‐density lipoprotein cholesterol, and low‐density lipoprotein cholesterol and higher serum concentrations of urea than the NM group. The partial replacement of alfalfa hay (≤50%) and maize silage with MO silage had no negative effects on milk yield, in vivo nutrient apparent digestibility or serum biochemical indexes of lactating cows.     

Prevalence and severity of foot lesions in Danish Holstein heifers through first lactation

Several studies have shown that foot lesions and clinical lameness occur before first calving and develop further during the lactation period. Lameness may cause production losses, but the relationship between foot lesions, particularly in the claw horn, and lameness in heifers is unclear. The objectives of this study were to describe the development of and evaluate the relationship between lameness and foot lesions in Danish Holsteins before and after first calving. In a longitudinal study, 147 heifers were examined for lameness and foot lesions 2–5 times over an 18-month period. Lameness was assessed by means of a visual locomotion score and foot lesion severity was recorded.The prevalence of a locomotion score 3 was 25% before calving, and 90% at approximately 250 days in milk (DIM). Prevalence of moderate to severe sole haemorrhage (SH) was 27% before calving and 56% at 250 DIM, and that of moderate to severe white line lesion (WLL) 44% before calving with a peak of 70% at 200 DIM. There was one case of white line abscess but SH was seen throughout the entire study period. Digital dermatitis (DD) was prevalent prior to first calving (15%) and peaked at 39% at 0–100 DIM. Heel horn erosion (HHE) occurred in almost all cows (93–100%) and was strongly correlated with DD (r = 0.51). The correlation coefficient between SH and WLL was also high (0.42). The relatively high correlations between WLL and both DD and HHE were more surprising (0.38 and 0.35, respectively), those between SH and both DD and HHE were moderate (around 0.18). Interdigital dermatitis was significantly correlated with both HHE and DD, but completely unrelated to SH and WLL.The overall average locomotion score increased by about one-half of a score unit from 1 month prior to calving until 250 DIM, with a large difference between herds, although this was unsurprising as cows may alter their locomotion pattern with management factors (e.g. floor properties). DD and WLL were both associated with a locomotion score 3 but of the cows with severe WLL there was no clear association between a locomotion score 3 and DD. The highest locomotion scores occurred among cows with DD but without WLL.     

Influence of calving difficulty on dry matter intake immediately after calving of dairy cows

We investigated the influence of calving difficulty on dry matter intake (DMI) in Holstein cows immediately after calving using 15 pregnant heifers and 15 multiparous cows. DMI was measured for 6 days after calving. Calving difficulty was evaluated with a calving score, and urinary cortisol was measured. The calving score was higher in the first lactation heifers than in the multiparous cows. The average DMI 6 days after calving was lower in the first lactation heifers than in the multiparous cows. The urinary cortisol concentration at 4 days after calving was higher in the first lactation heifers than in the multiparous cows and was positively associated with the calving score. The DMI was negatively associated with the calving score and the urinary cortisol concentration at 4 days after calving and was positively associated with the average milk yield 6 days after calving and the serum calcium concentration at 3 days after calving. We conclude that DMI immediately after calving is influenced not only by nutrient requirements of cows but also stress derived from calving, and the low DMI in the first lactation heifers is due to the lower energy requirement and high susceptibility to the stress compared with multiparous cows.     

Epidemiological investigation of Leptospira spp. in a dairy farming enterprise after the occurrence of three human leptospirosis cases

An epidemiological investigation was conducted in an unvaccinated dairy farming enterprise in which three workers on one of the milking herds (Herd 1) were diagnosed with leptospirosis due to serovars Hardjo (H) (n = 2) and Pomona (P) (n = 1) between January and March 2015. Blood and urine samples were collected from milking cows in Herd 1 (N = 230) and Herd 2 (N = 400), rising one‐ (R1, N = 125) and rising two‐year‐old (R2, N = 130) replacement heifers, and four pigs associated with Herd 1, in March 2015. Sera were tested using the MAT for serovars H, P, Copenhageni (C), Ballum (B) and Tarassovi (T), and urine samples were tested by qPCR. Seventy‐five per cent of 109 cows in Herd 1 and 36% of 121 in Herd 2 were seropositive (≥48), predominantly to H and P, and 23% of 74 cows in Herd 1 and 1% of 90 cows in Herd 2 were qPCR positive. Fifty‐five per cent of 42 R2 heifers were seropositive to T. No R1 and 17% of 42 R2 heifers were qPCR positive. Subsequently, all cattle were vaccinated for H and P, and Herds 1 and 2 were given amoxicillin. After the booster vaccination, 7% of 91 in Herd 1, 2% of 82 in Herd 2 and 11% of 38 R1 heifers (sampled as R2) were PCR positive. After the amoxicillin treatment, no cows in Herd 1 and 5% of 62 cows in Herd 2 were urine PCR positive. Calves and pigs were seropositive to H, P, C and B. Vaccination and antibiotic treatment appeared effective in reducing the risk of exposure of workers to vaccine serovars. However, evidence of non‐vaccine serovars indicated that workers likely remain at risk of exposure to Leptospira.     

Seroprevalence of Bartonella species,Coxiella burnetii and Toxoplasma gondii among patients with hematological malignancies: A pilot study in Romania

Patients receiving immunosuppressive cancer treatments in settings where there is a high degree of human–animal interaction may be at increased risk for opportunistic zoonotic infections or reactivation of latent infections. We sought to determine the seroprevalence of selected zoonotic pathogens among patients diagnosed with haematologic malignancies and undergoing chemotherapeutic treatments in Romania, where much of the general population lives and/or works in contact with livestock. A convenience sample of 51 patients with haematologic cancer undergoing chemotherapy at a referral clinic in Cluj‐Napoca, Romania, was surveyed regarding animal exposures. Blood samples were obtained and tested for evidence of infection with Bartonella species, Coxiella burnetii and Toxoplasma gondii, which are important opportunistic zoonotic agents in immunocompromised individuals. 58.8% of participants reported living or working on a farm, and living or working on a farm was associated with contact with livestock and other animals. 37.5% of participants were IgG seroreactive against one or more of five Bartonella antigens, and seroreactivity was statistically associated with living on farms. Farm dwellers were 3.6 times more likely to test IgG seroreactive to Bartonella antibodies than non‐farm dwellers. 47.1% of the participants tested T. gondii IgG positive and 13.7% tested C. burnetii IgG positive, indicating past or latent infection. C. burnetii IgM antibodies were detected in four participants (7.8%), indicating possible recent infection. These results indicate that a large proportion of patients with haematologic cancer in Romania may be at risk for zoonotic infections or for reactivation of latent zoonotic infections, particularly with respect to Bartonella species. Special attention should be paid to cancer patients' exposure to livestock and companion animals in areas where much of the population lives in rural settings.     

Early lactation dairy cows: Development of equations to predict intake and milk performance at grazing

The objectives of this study were to examine the effect of parity and days in milk (DIM) on dry matter intake (DMI) and actual milk yield (MY_Act) of grazing spring calving dairy cows in early lactation (< 100 DIM) and to develop equations to predict DMI and milk yield for grass based systems of milk production. A dataset containing 335 observations from 134 Holstein Friesian dairy cows was assembled from two early lactation grazing studies. Observations were available for primiparous (n = 130) and multiparous (n = 205) cows during periods of DMI measurement using the n-alkane technique. Animal performance was divided into two classes of DIM: less than 50 DIM (< 50 DIM) or between 51 and 100 DIM (> 50 DIM). Parity and DIM had a significant effect on grass DMI (GDMI), total DMI (TDMI), MY_Act and milk composition. TDMI increased with parity and DIM and ranged from 13.4 kg/cow per day (primiparous animals, < 50 DIM) to 20.1 kg/cow per day (multiparous animals, > 50 DIM). Actual MY increased with parity and decreased with DIM (range: 24.1 kg/cow per day (primiparous animals, > 50 DIM) to 33.0 kg/cow per day (multiparous animals, < 50 DIM)). Multiparous cows had greater bodyweight (BW) and lower BCS than primiparous cows. In the early lactation period a number of variables had a significant effect on GDMI, TDMI and milk yield. These predictor variables included BW, BCS, potential milk yield (MY_Pot), DIM, daily herbage allowance (DHA; > 4 cm), concentrate level and parity. The equations accounted for 79%, 83% and 86% of the variation in GDMI, TDMI and milk yield, respectively. Actual milk yield was always below the MY_Pot of the cows, the mean difference was 5.8 kg/cow per day. As DHA and concentrate level increased, the difference between MY_Act and MY_Pot reduced. This study supports the concept that immediately post-calving offering a grass based diet with a medium level of concentrate supplementation is sufficient to support high milk production in grazing dairy cows.     

Progression of Coxiella burnetii infection after implementing a two-year vaccination program in a naturally infected dairy cattle herd

Background

The high prevalence of Coxiella burnetii infection in dairy cattle herds recently reported and the long survival time of the bacterium in the environment pose a risk to human and animal health that calls for the implementation of control measures at herd level. This study presents the results of a 2-year vaccination program with an inactivated phase I vaccine in a Spanish dairy herd naturally infected with C. burnetii. Calves older than 3 months and non-pregnant heifers and cows were vaccinated in April 2011 and the farm was subsequently visited at a monthly basis for vaccination of recently calved cows and calves that reached the age of 3 months. Annual booster doses were given to previous vaccinated animals as well. The effectiveness of the vaccine was assessed in terms of level of C. burnetii shedding through milk and uterine fluids and environmental contamination as determined by polymerase chain reaction (PCR).

Results

The percentage of shedder animals through uterine fluids and milk progressively decreased, and C. burnetii DNA load in bulk-tank milk samples was low at the end of the study. The average seroconversion rate in not yet vaccinated animals, which acted as control group, was 8.6% during the first year and 0% in the second year. DNA of C. burnetii was found in aerosols and dust samples taken in the calving area only at the beginning of the study, whereas slurry samples remained C. burnetii PCR positive for at least 18 months. Multiple Locus Variable number tandem-repeat Analysis identified the same genotype in all C. burnetii DNA positive samples.

Conclusions

In the absence of any changes in biosecurity, the overall reduction of C. burnetii infection in animals to 1.2% milk shedders and the reduced environment contamination found at the end of the study was ascribed to the effects of vaccination together with the culling of milk shedders. Vaccination has to be planned as a medium-long term strategy to suppress risks of re-infection.     

Occupational swine exposure and Hepatitis E virus,Leptospira, Ascaris suum seropositivity and MRSA colonization in Austrian veterinarians, 2017–2018—A cross‐sectional study

We investigated the prevalence of Hepatitis E Virus (HEV), Leptospira and Ascaris suum (A. suum) seropositivity, and of nasal methicillin‐resistant Staphylococcus aureus (MRSA) colonization among Austrian practising veterinarians, and assessed the association with occupational swine livestock exposure. The 261 participants completed a questionnaire on demographics, intensity of occupational swine livestock contact and glove use during handling animals and their secretions. Participants' blood samples were tested for HEV, Leptospira and A. suum seropositivity and nasal swabs cultured for MRSA. We compared swine veterinarians (defined as >3 swine livestock visits/week) to non‐swine veterinarians (≤3 swine livestock visits/week) with regard to the outcomes through calculating prevalence ratio (PR) and 95% confidence interval (CI). Furthermore, the relationship between occupational swine livestock contact and the study outcomes was examined by age (</≥55 years) and glove usage. The prevalence of nasal MRSA colonization was 13.4% (95% CI: 9.3–17.6), of HEV seropositivity 20.8% (95% CI: 15.8–25.7) and A. suum seropositivity 44% (95% CI: 37.7–50.2). The highest anti‐leptospiral antibodies titres were 1:200 (L. hebdomadis) and 1:100 (L. autumnalis, L. caicola) found in three non‐swine veterinarians. Compared to non‐swine veterinarians, swine veterinarians were 1.9 (95% CI: 1.0–3.4) and 1.5 (95%CI: 1.0–2.3) times more likely HEV seropositive and A. suum seropositive, respectively, and 4.8 (95%CI: 2.5; 9.3) times more likely nasally colonized with MRSA. Among glove‐using veterinarians, occupational swine contact was no longer a determinant for HEV seropositivity (PR 1.6; 95% CI: 0.8–2.9). Similar was found for A. suum seropositivity, which was no longer associated with occupational swine livestock contact in the subgroup of glove using, ≥55‐year‐old veterinarians (PR: 1.07; 95% CI: 0.4–3.3). Our findings indicate that >3 occupational swine livestock visits per week is associated with HEV and A. suum seropositivity and nasal MRSA colonization and that glove use may play a putative preventive role in acquiring HEV and A. suum. Further analytical epidemiological studies have to prove the causality of these associations.     

Coxiella burnetii seroprevalence in unvaccinated veterinary workers in Australia: Evidence to support Q fever vaccination

Q fever (caused by Coxiella burnetii) is a serious zoonotic disease that occurs almost worldwide. Occupational contact with animals increases the risk of exposure, and Q fever vaccination is recommended for veterinary workers in Australia. This study aimed to investigate C. burnetii seroprevalence among unvaccinated veterinary workers in Australia and determine factors associated with a positive serological result. During 2014 and 2015, convenience sampling at veterinary conferences and workplace vaccination clinics was undertaken. Participants completed a questionnaire and provided a blood sample for C. burnetii serology. Participants were predominantly veterinarians (77%), but veterinary support staff, animal scientists, and administration workers also participated. Blood samples (n = 192) were analysed by an immunofluorescence assay and considered positive where the phase I or phase II IgG titre was ≥1/50. Seroprevalence was 19% (36/192; 95% CI 14%–25%). A positive serological result was significantly associated with (a) working in outer regional/remote areas (odds ratio [OR] 6.2; 95% CI 1.9–20.8; reference = major cities; p = .009) and (b) having spent more than 50% of total career working with ruminants (OR 4.8; 95% CI 1.7–13.5; reference = <15% of career; p = .025). These findings confirm an increased risk of exposure to C. burnetii compared to the general population, providing new evidence to support Q fever vaccination of veterinary workers in Australia.     

Subclinical ketosis in dairy cows: prevalence and risk factors in grazing production system

The purpose of this study was to estimate the prevalence of Subclinical ketosis (SCK) between 4 and 19 days in milk (DIM) in a grazing production system and investigate the importance of potential risk factors for SCK. This cross‐sectional study was conducted in dairy cows (n = 107), which had more of two parities. The concentration of β‐hydroxybutyric (BHB) in blood was quantified through a hand‐held meter. Potential risk factors evaluated were calving interval (CI), milk yield in previous lactation, metritis, dystocia, calf sex (male), parity (≤3 vs. ≥4) and pre‐partum body condition score (BCS ≤ 3.5 vs. ≥3.75). Prevalence of SCK was 10.3% (95% CI 4.7–15) between 4 and 19 DIM. Risk factors identified were the occurrence of both metritis and pre‐partum BCS ≥ 3.75. Cows with metritis had 4.9 (95% CI 1.17–20.98) times more risk of developing SCK than cows without metritis. And the cows with pre‐partum BCS ≥ 3.75 had 5.25 (95% CI 1.32–21.11) times more risk of developing SCK than cows with pre‐partum BCS ≤ 3.5. Metritis could induce a lower feed intake and promote the development of SCK. High pre‐partum BCS could induce a greater mobilization of body reserves altering liver function and aggravating post‐partum NEB. The results are indicative of the expected prevalence of SCK in grazing production system. Factors associated could help to identify cattle at risk of SCK and improve the management of strategies to limit the effects.