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    J.C.M. Vernooij

    Coccidiosis is an economically important disease in chickens, caused by infection with Eimeria species parasites. Diagnosis of coccidiosis is frequently based on oocyst enumeration in pooled faecal samples or litter. In studies on... more
    Coccidiosis is an economically important disease in chickens, caused by infection with Eimeria species parasites. Diagnosis of coccidiosis is frequently based on oocyst enumeration in pooled faecal samples or litter. In studies on infection dynamics and for monitoring in the field, samples from individual chickens may be more appropriate as these support the determination of infection status of individual birds and more accurately reflect oocyst output at time of sampling. Faecal samples from individual birds can be collected, but the counting procedure limits the number of samples that can be processed and unequivocal microscopic differentiation between Eimeria species is very difficult. A test that overcomes these drawbacks would improve efficiency and quality of the diagnosis. The aim of this study was to compare two methods for Eimeria oocyst quantification in samples from individual birds. A real-time PCR that quantifies oocysts in cloacal swabs (qPCR) and oocyst counts in sing...
    Shortening or omitting the dry period (DP) has been proposed as a management strategy to improve energy balance of dairy cows in early lactation. Both shortening and complete omission of the DP reduces milk production in the subsequent... more
    Shortening or omitting the dry period (DP) has been proposed as a management strategy to improve energy balance of dairy cows in early lactation. Both shortening and complete omission of the DP reduces milk production in the subsequent lactation compared with a conventional DP length of 60d. Some cows have less milk production loss than other cows after applying no DP or a short DP. The aim of this study is to evaluate which cow characteristics are associated with the amount of milk production losses following no DP or a short DP (30d). Daily production information from the lactation before and after the DP was available from 161 dairy cows (54 cows with a 0-d DP, 51 cows with a 30-d DP, and 56 cows with a 60-d DP) from a research herd. Daily production (milk, fat, and protein) until 305d in milk was estimated for all cows. Subsequently, total fat- and protein-corrected milk yield from 60d before the expected calving date until 305d in the following lactation (FPCMtotal) was estimat...
    Staphylococcus aureus is one of the most prevalent causes of bovine mastitis. The antimicrobial treatment of this disease is currently based on antimicrobial susceptibility tests according to Clinical and Laboratory Standards Institute... more
    Staphylococcus aureus is one of the most prevalent causes of bovine mastitis. The antimicrobial treatment of this disease is currently based on antimicrobial susceptibility tests according to Clinical and Laboratory Standards Institute standards. However, various authors have shown a discrepancy between the results of this standard susceptibility test and the actual cure rate of the applied antimicrobial treatment. Increasing evidence suggests that in vivo biofilm formation by Staph. aureus, which is not assessed in the antimicrobial susceptibility tests, is associated with this problem, resulting in disappointing cure rates, especially for infections of longer duration. Previous data obtained with a limited number of strains showed that the extended biofilm antimicrobial susceptibility (EBS) assay reveals differences between strains, which cannot be derived from a standard susceptibility test or from a 24-h biofilm susceptibility test. The objective of this study was to test a coll...
    Before interventions to control horizontal transmission of Mycoplasma gallisepticum can be tested, a suitable experimental model should be available. Transmission dynamics in a flock can be quantified by two parameters: the average number... more
    Before interventions to control horizontal transmission of Mycoplasma gallisepticum can be tested, a suitable experimental model should be available. Transmission dynamics in a flock can be quantified by two parameters: the average number of secondary cases infected by one typical infectious case (R0) and the number of new infections that occur due to one infectious animal per unit of time (beta). The transmission dynamics of M. gallisepticum have not been studied experimentally, so the aim of this study was to examine the horizontal transmission of M. gallisepticum. The study was carried out using a pairwise design with three different inoculation doses. Every pair consisted of an inoculated chicken and a susceptible in-contact chicken. Five susceptible individually housed chickens were placed in between pairs in order to measure airborne transmission. Infection was detected by serology, quantitative polymerase chain reaction and culture. The inoculated and in-contact chickens were...
    Calves derived from IVP embryos may suffer from the large offspring syndrome that has been related to effects of in vitro culture on the intrinsic quality of the embryo. Limited information is available on the role of the placenta in such... more
    Calves derived from IVP embryos may suffer from the large offspring syndrome that has been related to effects of in vitro culture on the intrinsic quality of the embryo. Limited information is available on the role of the placenta in such cases. In this study, bovine pregnancy-associated glycoprotein (bPAG) was used as a marker to test whether placental function is influenced by the route of embryo production. Therefore, from day 7 until day 119 of ongoing gestations, resulting from transfer of MOET (n = 53), IVP-co-culture (n = 21) and IVP-SOF (n = 38) embryos, bPAG levels were compared in peripheral plasma of recipients. Plasma progesterone levels were compared as well. From day 25 of gestation onwards, bPAG could be detected in all recipients and the levels were significantly influenced by the day of gestation. Although IVP calves were significantly heavier than the in vivo produced calves, this difference was not reflected in the bPAG profiles of the embryo production groups. Ye...
    The objective of this research was to determine the contribution of cow factors to the probability of successful insemination accounting for the serial number of inseminations in analysis. The investigation was performed with 101,297... more
    The objective of this research was to determine the contribution of cow factors to the probability of successful insemination accounting for the serial number of inseminations in analysis. The investigation was performed with 101,297 insemination records in 51,525 lactations of different cows from 1368 herds obtained from the Dutch milk production recording database. Cows that had a first insemination (AI) between 40 and 150 days post-partum with one or more inseminations (≤6 inseminations) were selected. An insemination was defined successful when not followed by another insemination and when the cow calved between 267 and 295 days after insemination, or when the cow was culled between 135 and 295 days after the last insemination. Breed, parity, days in milk, lactation curve characteristics, milk production traits, moment of AI related to peak milk yield time (before or after peak milk yield), the last calf (female, male, twin or stillbirth) and season of insemination were selected as independent parameters for a model with successful rate of insemination as dependent parameter. A multivariable logistic regression model was used within cow and farm as a random effect. The probability of successful insemination was the highest in the first insemination and decreased in the following inseminations. However, the success rate of all inseminations increased in a later stage of lactation. The improvement in the successful inseminations in a later stage of lactation was better in multiparous cows than in first parity cows. Insemination in summer and before peak milk yield time reduced the success of insemination. The success rate was the lowest in 100% Holstein Friesian cows compared with other breeds and was the highest when the last calf was a female calf compared to a male calf, twin or stillbirth. In conclusion, the success of first and following inseminations depended on parity, breed, season of insemination, last calf status, daily milk yield at insemination date, serial insemination number and days in milk at insemination date.
    Resistance to coccidiostats and possible future restrictions on their use raise the need for alternative methods of reducing coccidiosis in poultry. The aim of this study was to evaluate the effect of selected phytochemicals on Eimeria... more
    Resistance to coccidiostats and possible future restrictions on their use raise the need for alternative methods of reducing coccidiosis in poultry. The aim of this study was to evaluate the effect of selected phytochemicals on Eimeria tenella sporozoite invasion in vitro. Four phytochemicals were selected on the basis that they reduce the virulence of Eimeria spp. and/or provide immune modulatory benefits to host cells: betaine, carvacrol, curcumin and Echinacea purpurea extract (EP). Madin-Darby bovine kidney (MDBK) cells were covered by medium containing phytochemicals at the highest concentration which was non-toxic to the cells. Salinomycin 50 μg/ml was positive control; negative control was medium only. E. tenella (Houghton strain) sporozoites were added to wells and after incubation for 2, 4 or 20 h at 37°C, cells were fixed and stained with hematoxylin-eosin. Ten evenly spaced fields per well were photographed and the percentage of cells invaded by sporozoites was calculated and normalized to the control. At 2h, carvacrol, curcumin and EP showed a significantly lower percentage of sporozoite invasion than the untreated control; in contrast, betaine treatment represented a significantly higher invasion percentage. Combining carvacrol with EP inhibited E. tenella invasion more effectively than applying the compounds individually, but the further addition of curcumin did not reduce invasion further. In conclusion, this study shows that invasion of MDBK epithelial cells by E. tenella sporozoites is inhibited in the presence of carvacrol, curcumin, or EP and enhanced by betaine. There may be potential for developing these phytochemicals as anti-coccidial feed or water additives for poultry.
    The aim of this study was to determine the association between farm management factors, including antimicrobial drug usage, and resistance in commensal Escherichia coli isolates from the faeces of white veal calves. Ninety E. coli... more
    The aim of this study was to determine the association between farm management factors, including antimicrobial drug usage, and resistance in commensal Escherichia coli isolates from the faeces of white veal calves. Ninety E. coli isolates from one pooled sample per farm (n = 48) were tested for their phenotypical resistance against amoxicillin, tetracycline, cefotaxime, ciprofloxacin and trimethoprim/sulfamethoxazole (TMP/SMX). Logistic regression analysis revealed the following risk factors (P < 0·05); farmer wearing the same work clothes for several days [ciprofloxacin, odds ratio (OR) 2·6; tetracycline, OR 2·4], administration of trimethoprim-sulfonamide combinations (TMP/SMX, OR 3·0; amoxicillin, OR 3·1; tetracycline, OR 2·6), ⩾0·3 animal daily dosage per production cycle (ADD/pc), quinolones (ciprofloxacin, OR 2·8), ⩾1·3 ADD/pc, penicillins (ciprofloxacin, OR 3·3; tetracycline, OR 3·4), 20-40 ADD/pc, tetracyclines (tetracycline, OR 3·2) and >40 ADD/pc, tetracyclines (tetracycline, OR 13·1; amoxicillin, OR 6·5). In this study antimicrobial resistance in commensal E. coli was mainly associated with antimicrobial drug use.
    The effect of an inactivated vaccine on the horizontal transmission of Mycoplasma gallisepticum was quantified in a transmission model. Twenty non-vaccinated and 20 vaccinated 23-week-old specific pathogen free hens were housed in pairs,... more
    The effect of an inactivated vaccine on the horizontal transmission of Mycoplasma gallisepticum was quantified in a transmission model. Twenty non-vaccinated and 20 vaccinated 23-week-old specific pathogen free hens were housed in pairs, while five individually housed hens acted as a negative control group. Each pair consisted of a challenged chicken (10(4) colony forming units intratracheally) and a non-challenged susceptible contact bird. Infection was monitored by serology, quantitative polymerase chain reaction and culture. All non-vaccinated and vaccinated in-contact chickens became infected with M. gallisepticum. The 95% confidence interval of the reproduction ratio, R (a measure of transmission defined as the average number of secondary cases caused by one infectious individual) was 4.48 to infinity in both groups. However, the logarithm of the area under the curve in the vaccinated group was 0.51 lower (P = 0.02) than in the non-vaccinated group, indicating that there was an effect of vaccination on the levels of potential shedding of M. gallisepticum. Nevertheless, the results of this study indicate that the use of an inactivated M. gallisepticum vaccine will not reduce the horizontal transmission of M. gallisepticum between laying hens.
    The effect of a live Mycoplasma gallisepticum vaccine on the horizontal transmission of this Mycoplasma species was quantified in an experimental animal transmission model in specific pathogen free White Layers. Two identical trials were... more
    The effect of a live Mycoplasma gallisepticum vaccine on the horizontal transmission of this Mycoplasma species was quantified in an experimental animal transmission model in specific pathogen free White Layers. Two identical trials were performed, each consisting of two experimental groups and one control group. The experimental groups each consisted of 20 birds 21 weeks of age, which were housed following a pair-wise design. One group was vaccinated twice with a commercially available live attenuated M. gallisepticum vaccine, while the other group was not vaccinated. Each pair of the experimental group consisted of a challenged chicken (10(4) colony-forming units intratracheally) and a susceptible in-contact bird. The control group consisted of 10 twice-vaccinated birds housed in pairs and five individually housed non-vaccinated birds. The infection was monitored by serology, culture and quantitative polymerase chain reaction. The vaccine strain and the challenge strain were distinguished by a specific polymerase chain reaction and by random amplified polymorphic DNA analysis. In both experiments, all non-vaccinated challenged chickens and their in-contact 'partners' became infected with M. gallisepticum. In the vaccinated challenged and corresponding in-contact birds, a total of 19 and 13 chickens, respectively, became infected with M. gallisepticum. Analysis of the M. gallisepticum shedding patterns showed a significant effect of vaccination on the shedding levels of the vaccinated in-contact chickens. Moreover, the Cox Proportional Hazard analysis indicated that the rate of M. gallisepticum transmission from challenged to in-contact birds in the vaccinated group was 0.356 times that of the non-vaccinated group. In addition, the overall estimate of R (the average number of secondary cases infected by one typical infectious case) of the vaccinated group (R = 4.3, 95% confidence interval = 1.6 to 49.9) was significantly lower than that of the non-vaccinated group (R = infinity, 95% confidence interval = 9.9 to infinity). However, the overall estimate of R in the vaccinated group still exceeded 1, which indicates that the effect of the vaccination on the horizontal transmission M. gallisepticum is insufficient to stop its spread under these experimental conditions.
    The notochordal cell (NC) of the nucleus pulposus (NP) is considered a potential NP progenitor cell, and early interverte-bral disk (IVD) degeneration involves replacement of NCs by chondrocyte-like cells (CLCs). Wnt/b-catenin signaling... more
    The notochordal cell (NC) of the nucleus pulposus (NP) is considered a potential NP progenitor cell, and early interverte-bral disk (IVD) degeneration involves replacement of NCs by chondrocyte-like cells (CLCs). Wnt/b-catenin signaling plays a crucial role in maintaining the notochordal fate during embryogenesis, but is also involved in tissue degeneration and regeneration. The canine species, which can be subdivided into non-chondrodystrophic and chondrodystrophic breeds, is characterized by differential maintenance of the NC: in non-chondrodystrophic dogs, the NC remains the predominant cell type during the majority of life, with IVD degeneration only occurring at old age; conversely, in chondrodystrophic dogs the NC is lost early in life, with concurrent degeneration of all IVDs. This study investigated Wnt/b-catenin signaling in the healthy, NC-rich NP and early degenerated, CLC-rich NP of both breed types by immunohistochemistry of b-catenin and relative gene expression of brachyury and cytokeratin 8 (notochordal markers) and Wnt targets axin2, cyclin D1, and c-myc. Both NCs and CLCs showed nuclear and cytoplasmic b-catenin protein expression and axin2 gene expression, but b-catenin signal intensity and Wnt target gene expression were higher in the CLC-rich NP. Primary NCs in monolayer culture (normoxic conditions) showed Wnt/b-catenin signaling comparable to the in vivo situation, with increased cyclin D1 and c-myc gene expression. In conclusion, Wnt/b-catenin signaling activity in the NC within the NC-rich NP and in culture supports the role of this cell as a potential progenitor cell; increased Wnt/b-catenin signaling activity in early IVD degeneration may be a reflection of its dual role.
    Medial coronoid disease (MCD) is a common joint disease of dogs. It has a multifactorial aetiology, but the relationship between known causal factors and the disease has yet to be elucidated. As most of the published literature is... more
    Medial coronoid disease (MCD) is a common joint disease of dogs. It has a multifactorial aetiology, but the relationship between known causal factors and the disease has yet to be elucidated. As most of the published literature is clinical and it reports changes associated with advanced disease, it is not known whether the changes reflect the cause or consequences of the condition. The aim of this study was to investigate early micromorphological changes occurring in articular cartilage and to describe the postnatal development of the medial coronoid process (MCP) before MCD develops. Three litters of MCD-prone young Labrador retrievers were purpose-bred from a dam and two sires with MCD. Comparisons of the micromorphological appearance of the MCP in MCD-negative and MCD-positive joints demonstrated that MCD was initially associated with a disturbance of endochondral ossification, namely a delay in the calcification of the calcifying zone, without concurrent abnormalities in the superficial layers of the joint cartilage. Cartilage canals containing patent blood vessels were only detected in dogs <12 weeks old, but the role of these channels in impaired ossification requires further investigation. Retained hyaline cartilage might ossify as the disease progresses, but weak areas can develop into cracks between the retained cartilage and the subchondral bone, leading to cleft formation and fragmentation of the MCP.