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Neonatal mortality in puppies is a problem frequently encountered by dog breeders. Often, only postmortem examination allows diagnosis and implementation of measures to save the rest of the litter. This article presents the key steps of the postmortem examination, namely, autopsy, histopathology, bacteriology, molecular identification of pathogens, and coproscopy. Sampling, samples' conservation, and interpretation of the obtained results are presented as well as their relative importance for the final diagnosis. Finally, examples of the most frequent syndromes observed under postmortem examination in canine newborns, together with the results from a complementary analysis looking for infectious agents responsible for death, are discussed.

Neonatal mortality in puppies is a problem frequently encountered by dog breeders. Often, only postmortem examination allows diagnosis and implementation of measures to save the rest of the litter. This article presents the key steps of the postmortem examination, namely, autopsy, histopathology, bacteriology, molecular identification of pathogens, and coproscopy. Sampling, samples' conservation, and interpretation of the obtained results are presented as well as their relative importance for the final diagnosis. Finally, examples of the most frequent syndromes observed under postmortem examination in canine newborns, together with the results from a complementary analysis looking for infectious agents responsible for death, are discussed.

Bisphenol S (BPS) is widely used as a substitute for Bisphenol A in consumer products. Despite its potential endocrine-disrupting effects and widespread exposure, toxicokinetic data, particularly during the critical period of pregnancy, are not available for BPS. The objectives of our study were to evaluate the mechanisms determining fetal exposure to BPS and to BPS glucuronide (BPSG) and to compare them with those prevailing for BPA.The disposition of BPS and BPSG was evaluated in the materno-fetal unit of the catheterized pregnant ewe model, following intravenous administrations of BPS and BPSG to mothers and their fetuses. In a second experiment, the rate of BPS accumulation in the fetal compartment was determined under steady-state conditions after repeated intravenous BPS administrations to the mother.In the maternal compartment, BPS was mainly metabolized into BPSG and totally eliminated in urine. Only 0.40% of the maternal dose was transferred to the fetus. However, once in the fetal compartment, 26% of the fetal dose was rapidly eliminated through placental transfer, while 46% of BPS was metabolized into BPSG which remained trapped in the fetal compartment. Thus, the elimination of BPSG from the fetal compartment required its back-conversion into bioactive BPS, leading to an 87% enhancement of the fetal BPS exposure.Our findings demonstrate that, despite the low materno-fetal placental transfer of BPS, this substitute for BPA is able to accumulate in the fetal compartment after repeated maternal exposure, leading to chronic fetal exposure to BPS in a range of concentrations similar to those of BPA. Keywords: Bisphenol S, Bisphenol S Glucuronide, Toxicokinetic, Fetal exposure, Bisphenol A analogue

Highlights • As in farm animals, deficit in passive immune transfer is associated in puppies with a higher risk of neonatal mortality. • Intestinal barrier is closed after 12–16 hours of life in puppies. • Time of colostrum ingestion is the major determinant of passive immune transfer, rather than colostral IgG concentration. • Banking of colostrum collected between 24 and 48 hours of life should be encouraged. • Lactogenic immune transfer is not only systemic but also local, providing protection of the digestive tract., The puppy, born without immunoglobulins G (IgG), acquires a passive systemic immunity thanks to colostrum intake during the two first days of life. The quality of passive immune transfer (i.e. blood IgG concentration at two days of age), highly variable between litters and between puppies within litters, depends mainly on the time elapsed between birth and ingestion of colostrum, with limited influence of colostrum IgG concentration. Deficit in passive immune transfer, impacting puppy’s health and neonatal mortality rate, can be indirectly diagnosed through blood gammaglutamyltransferases assay and evaluation of growth rate over the two first days of life. In the absence of maternal colostrum, few homo- and heterospecific immune sources are available and canine colostrum banking remains the optimal solution. Whereas passive immune transfer is crucial for survival during the neonatal period, it later interferes with response to vaccination. In addition to systemic passive immune transfer, maternal antibodies (mainly IgA) would provide local (digestive) immunity, ensuring mid-term protection of the puppies’ gut together with probably long term training of the digestive immune system.

Background: Given its hormonal activity, bisphenol S (BPS) as a substitute for bisphenol A (BPA) could actually increase the risk of endocrine disruption if its toxicokinetic (TK) properties, namely its oral availability and systemic persistency, were higher than those of BPA. Objectives: The TK behavior of BPA and BPS was investigated by administering the two compounds by intravenous and oral routes in piglet, a known valid model for investigating oral TK. Methods: Experiments were conducted in piglets to evaluate the kinetics of BPA, BPS, and their glucuronoconjugated metabolites in plasma and urine after intravenous administration of BPA, BPS, and BPS glucuronide (BPSG) and gavage administration of BPA and BPS. A population semiphysiologically based TK model describing the disposition of BPA and BPS and their glucuronides was built from these data to estimate the key TK parameters that drive the internal exposure to active compounds. Results: The data indicated that almost all the BPS oral dose was absorbed and transported into the liver where only 41% of BPS was glucuronidated, leading to a systemic bioavailability of 57.4%. In contrast, only 77% of the oral dose of BPA was absorbed and underwent an extensive first-pass glucuronidation either in the gut (44%) or in the liver (53%), thus accounting for the low systemic bioavailability of BPA (0.50%). Due to the higher systemic availability of BPS, in comparison with BPA, and its lower plasma clearance (3.5 times lower), the oral BPS systemic exposure was on average about 250 times higher than for BPA for an equal oral molar dose of the two compounds. Conclusion: Given the similar digestive tracts of pigs and humans, our results suggest that replacing BPA with BPS will likely lead to increased internal exposure to an endocrine-active compound that would be of concern for human health. https://doi.org/10.1289/EHP4599

International audience; Abstract: In numerous species, low birth weight is a risk factor for neonatal mortality. In the canine species, definition of a low birth weight is complex due to the huge interbreed variability in size. To identify puppies at higher risk of neonatal death, data from 6,694 puppies were analysed. The data were collected from 75 French breeding kennels, examining 27 breeds and totaling 1,202 litters of puppies. Generalised linear mixed models allowed to identify birth weight, birth weight heterogeneity within the litter, and size of the breeding kennel as significant risk factors for neonatal mortality. Receiver Operating Characteristics (ROC) and classification and regression tree (CART) analyses were combined to define breed specific thresholds for birth weight allowing the identification of puppies at higher risk of neonatal mortality. Due to differences in birth weights between breeds, including when belonging to the same breed size, analyses were conducted at the breed level. First, ROC analysis thresholds were successfully established for 12 breeds (area under the ROC ≥ 0.70; sensitivity ≥ 75%; specificity: 45–68%) and they ranged from 162 g in the Maltese to 480 g in the Bernese Mountain dog. Secondly, CART analysis thresholds from 22 breeds ranged from 105 g in the Maltese and 436 g in the Boxer. Puppies were grouped into three categories according to birth weight: low, moderate and high risk of neonatal mortality (higher than the ROC threshold, between ROC and CART thresholds, and lower than the CART threshold respectively). In the current study, 44% of the puppies were classified as at moderate risk and 5.3% for a high risk of neonatal mortality. Thresholds defined by CART analysis (and not ROC analysis) were used to define low birth weight puppies and were sometimes quite different between breeds with similar birth weight distributions suggesting a variable relationship between birth weight reduction and neonatal death. These results allow the identification of puppies at an increased risk of neonatal death, thus requiring specific nursing to improve their chances of survival. With these high risk puppies identified, both animal welfare and kennel productivity is predicted to improve.

International audience; The aim of our study was to evaluate the bidirectional transfer of Bisphenol S (BPS) and its main metabolite, BPS Glucuronide (BPSG), using the model of perfused human placenta and to compare the obtained values with those of Bisphenol A (BPA) and BPA Glucuronide. Fourteen placentas at term were perfused in an open dual circuit with deuterated BPS (1 and 5 μM) and non-labelled BPSG (2.5 μM) and a freely diffusing marker antipyrine (800 ng/ml) in the presence of albumin (25 mg/ml). In a second experiment, the potential role of P-glycoprotein in the active efflux of BPS across the placental barrier was studied using the well-established P-glycoprotein inhibitor, PSC833 (2 and 4 μM). Placental transfer of BPS was much lower than that of BPA in both directions. The placental clearance index of BPS in the materno-fetal direction was three times lower than in the opposite direction, strongly suggesting some active efflux transport. However, our results show that P-glycoprotein is not involved in limiting the materno-fetal transfer of BPS. Placental transfer of BPSG in the fetal compartment was almost non-existent indicating that, in the fetal compartment, BPSG originates mainly from feto-placental metabolism. The feto-maternal clearance index for BPSG was 20-fold higher than the materno-fetal index. We conclude that the blood-placental barrier is much more efficient in limiting fetal exposure to BPS than to BPA, indicating that the placenta has a crucial role in protecting the human fetus from BPS exposure.

International audience; Contents Failure of passive immune transfer put puppies at a higher risk of neonatal and weaning mortality due to low immune protection against infectious agents. The aim of this study was to investigate the evolution of the general via serum IgG concentration (IgG) and the specific via serum maternally derived canine parvovirus type 2-specific antibody titer (CPV2 MDA) passive immune transfer within the first 4 weeks of age. Furthermore, the relationship between general and specific immune transfer and the possibility of non-invasive evaluation was assessed. Puppies (169) were weighed systematically between birth and Day 28. IgG and CPV2 MDA were assayed in serum samples at 2 and at 28 days of age. At Day 2, there was a positive correlation between IgG and CPV2 MDA (rho = 0.71; p < 0.001). At Day 2, 17.9% (27/151) of puppies presented a deficit of passive immune transfer according to IgG result (defined as IgG < 2.3 g/L) and 25.8% (39/151) of puppies were under the minimal protective CPV2 MDA titer (defined as

Breeding kennels face a high rate of neonatal mortality, on which the impact of nutrition remains to be determined. This study was designed to evaluate the impact of birth weight (reflecting intrauterine growth) and early growth rate (reflecting colostrum intake) on risk of neonatal mortality in puppies and to determine the critical thresholds of both parameters. Puppies from various breeds were weighed at birth ( = 514) and at 2 d of age, and the growth rate over that period (early growth rate) was calculated for all survivors ( = 477). Linear mixed models evaluated the effect of birth weight on mortality between birth and 2 d of age and the effect of both birth weight and early growth rate on mortality between 2 and 21 d of age. Birth weight was influenced by litter size ( = 0.003), with more low-birth-weight puppies (the lightest 25% within a breed size) in large litters compared with smaller litters. Mortality over the first 2 d after birth was associated with birth weight ( < 0.001), with 81.1% of dying puppies characterized by a low birth weight. Mortality between 2 and 21 d of age was not related to birth weight but was found to be associated with early growth rate ( < 0.001), with higher risk of death in puppies with growth rate at or below -4% after the first 2 d of life. This study demonstrates the differential effect of intrauterine nutrition impacting mortality during the first 2 d of life and that of colostrum intake impacting mortality until 21 d of life. Birth weight and early growth rate thresholds provided in this study allow identification of puppies at risk, whereby provision can be made for adequate nursing to increase their chances to survive.

International audience

International audience

International audience; The prevalence of neonatal mortality is high in the canine species and far from well-studied. In most domestic neonates, an appropriate colostrum intake is a key element of the control of neonatal mortality. The aim of this study was to evaluate the impact of passive immune transfer on puppy mortality, assessed through serum immunoglobulin G (IgG) concentration at 2 days of age. Factors impacting passive immune transfer and the value of an oral immunoglobulin supplementation to prevent it were also analyzed. A total of 149 puppies from 34 litters (12 breeds) within one breeding kennel were included. Blood samples were collected at 2 days of age and colostrum was collected from their dams 1 day after whelping to assay IgG concentration. Puppies were weighed at birth and at 2 days of age for calculation of growth rate. Mortality was recorded until 3 weeks of age. Seventy randomly assigned puppies were orally supplemented with hyper-immunized adult plasma twice within the first 8h of life. IgG concentration at 2 days of age was significantly correlated with weight gain during the first 2 days of life. The multivariable model with litter as a random effect demonstrated that neonatal mortality was not influenced by breed size, sex, supplementation, litter size, nor colostrum IgG concentration, but by puppy IgG concentration at 2 days of age. According to the ROC curve, the minimal IgG concentration at and below which puppies were at higher risk of death was determined at 230mg/dl. Puppy IgG concentration was significantly associated with growth rate, but not with breed size, sex, supplementation, litter size or colostrum IgG concentration in a multivariable model with litter as a random effect. This study demonstrates that neonatal mortality in puppies is related to the quality of passive immune transfer. The oral supplementation with hyper-immunized canine plasma neither decreased risk of mortality, nor improved serum IgG concentration at 2 days of age in puppies. Attention must thus be paid to early colostrum intake to control the neonatal mortality in puppies.

The objective of this study was to describe efficiency of reproduction of purebred dogs in field breeding conditions, from mating to weaning in France. Data were collected between 2010 and 2014 in 5,667 French breeding kennels via a reproduction management software (Breeding Management System, Royal Canin, Aimargues, France). Effect of breed size (Mini: adult body weight 40 kg), age of dam and male on pregnancy rate, abortion rate and litter size were evaluated by multivariable models. Data on 45,913 heats (all with mating), from 27,221 bitches from 248 breeds, were analysed. At mating, mean age (±SD) was 3.1 ± 1.8 years for bitches and 3.3 ± 2.0 for males. Males originated from the same kennel as the females in 88.5% of the matings. Based on breeder's evaluation of the pregnancy status, pregnancy rate (number of pregnant females based on breeders declaration/number of heats) was 87.8% and abortion rate was 6.8%. Finally, 81.9% of the mated females gave birth to a litter. On 37,946 litters (204,537 puppies), mean litter size was 5.4 ± 2.8 puppies (range 1-24), which was influenced by breed size and dam age (p

Contents Colostrum and milk are complex mammary secretions providing the puppy with many nutritional and immunological factors, which play a crucial role for its correct development and survival. In the case of colostrum and/or milk intake deficiency, puppies are at increased risk of infectious diseases. This work reviews the various nutritional hyperimmune supplementations proposed to provide a passive immune protection and to positively impact puppies’ health. Some strategies rely on canine immunoglobulins: canine colostrum banking and canine serum/plasma supplementation. Others involve heterologous sources of antibodies and other immune factors: bovine colostrum or hyperimmune egg powder. Among the different solutions evaluated from birth to weaning, canine plasma and hyperimmune egg powder showed promising beneficial effect on puppies’ health. Canine plasma seems to positively impact not only growth (increased growth during the neonatal period), but also digestive health (higher species richness of intestinal microbiota) and the general health (tendency of lower morbidity). Puppies supplemented with hyperimmune egg powder presented increased neonatal growth and decreased risk of canine parvovirus infection. Nevertheless, natural canine maternal colostrum and milk ingestion remains the optimal guarantee for puppies’ health and survival, as a source of immunity, energy and growth factors.

Reproduction management and performances are evaluated in the feline species only through a limited number of animals and studies. Our objective was to provide reference figures in purebred cats, from a large-scale sample. Data were collected from an online software dedicated to cattery management (Breeding Management System®, BMS, Royal Canin, Aimargues, France). Information was recorded on a voluntary basis by French breeders between 2011 and 2014. Data were anonymously transferred for analysis. A total of 9,063 oestrous periods (in contact with a male) from 5,303 queens (45 breeds) were recorded from 1,521 breeders. Most matings (70.1%) occurred during increasing day length periods. The mean age at mating (±SD) was 2.7 ± 1.6 years for queens and 2.9 ± 1.9 years for tomcats. Pregnancy rate (based on breeders declaration) was 85.2%. Among queens declared pregnant, 8.4% failed to maintain pregnancy. Globally, 78% of the mated females gave birth to 28,065 kittens within 7,075 L. Mean litter size was 4.0 ± 1.9 kittens among which 8.5% were stillborn. Neonatal and paediatric mortality rate was 8.2%. In total, 16.0% of kittens born died before weaning. The results of this study are based on the largest feline database ever analysed. The figures collected can thus be used as reference to define average reproductive performances in numerous breeds for cat breeders. Further analysis will identify factors influencing reproductive performances and early mortality in the feline species.

Puppy and kitten survival over the first weeks is particularly dependent on colostrum, a specific secretion of the mammary gland produced during the first 2 days post-partum. Colostrum is a source of nutrients and immunoglobulins. It also contributes to the digestive tract maturation. Colostrum differentiates from milk mainly based on its concentration in immunoglobulins G: 20-30 g/L in dog colostrum, 40-50 g/L in cats' vs

Despite the high neonatal mortality rate in puppies, pertinent criteria for health evaluation of the newborns are not defined. This study was thus designed to measure and to characterize factors of variation of six health parameters in dog neonates, and to evaluate their value as predictors of neonatal mortality. A total of 347 purebred puppies under identical conditions of housing and management were examined within the first 8h after birth and then at Day 1. The first health evaluation included Apgar score, weight, blood glucose, lactate and β-hydroxybutyrate concentration, rectal temperature and urine specific gravity (SG). The second evaluation at Day 1 included the same parameters, excluding Apgar score and weight. The mortality rate over the first 24h and over 21days of age was recorded. The early predictors of neonatal mortality in the dog were determined with generalized linear mixed models and receiver operating characteristic curves analyses. An Apgar score at or below 6 evaluated within the first 8h after birth was found associated with a higher risk of death during the first 24h. A reduced glucose concentration (≤92mg/dl) at Day 1 was found to be associated with higher mortality between 1 and 21days of age. Low-birth-weight puppies were characterized by both low viability (low Apgar score) and low blood glucose concentration, and thus were found indirectly at higher risk of neonatal mortality. This study promotes two low cost easy-to-use tests for health evaluation in puppies, i.e. Apgar scoring and blood glucose assay. Further investigation is necessary to establish if the strong relationship between blood glucose and neonatal survival reflects high energy requirements or other benefits from colostrum intake.

International audience; Limited information is available describing the development of the neonatal fecal microbiome in dogs. Feces from puppies were collected at 2, 21, 42, and 56 days after birth. Feces were also collected from the puppies’ mothers at a single time point within 24 hours after parturition. DNA was extracted from fecal samples and 454-pyrosequencing was used to profile 16S rRNA genes. Species richness continued to increase significantly from 2 days of age until 42 days of age in puppies. Furthermore, microbial communities clustered separately from each other at 2, 21, and 42 days of age. The microbial communities belonging to dams clustered separately from that of puppies at any given time point. Major phylogenetic changes were noted at all taxonomic levels with the most profound changes being a shift from primarily Firmicutes in puppies at 2 days of age to a co-dominance of Bacteroidetes, Fusobacteria, and Firmicutes by 21 days of age. Further studies are needed to elucidate the relationship between puppy microbiota development, physiological growth, neonatal survival, and morbidity.

International audience; Canine neonates are born hypogammaglobulinemic, and colostrum is their main source of immunoglobulins. The purpose of this study was to evaluate the immune quality of canine colostrum and its variability both among bitches and among mammary glands. The immune quality was estimated from immunoglobulin G (IgG) concentration (ELISA test). The correlation of IgG concentration with refractometry was evaluated. From a total of 44 bitches from 13 different breeds from a single breeding kennel, samples of colostrum and blood were collected one day after the parturition onset. Colostrum was collected separately from each pair of mammary glands (180 pairs). The mean colostrum IgG concentration in our population was 20.8 +/- 8.1 g/L (ranging from 8.0 to 41.7 g/L) with no influence of breed size, litter size, age of dam or serum IgG concentration. Colostrum IgG concentration varied widely among pairs of mammary glands within one bitch (variation coefficient: 42 +/- 32.1%). Nevertheless, no single pair of mammary glands was found to produce regularly a secretion of higher quality. No difference in IgG concentration was recorded between anterior and posterior pairs either. The BRIX index and the refractive index were significantly, but moderately correlated with colostrum IgG concentration (r=0.53 and 0.42, respectively). This study demonstrates a great variability in immune quality of colostrum among bitches and among mammary glands within one bitch. Further studies on the suckling behavior of puppies and on determination of the minimal immune quality of colostrum are required to evaluate their impact of this high variability on neonatal mortality in dogs. (C) 2015 The Authors. Published by Elsevier Ireland Ltd.