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Animals, Newborn - Top 30 Publications

Impact of maternal high fat diet on hypothalamic transcriptome in neonatal Sprague Dawley rats.

Maternal consumption of a high fat diet during early development has been shown to impact the formation of hypothalamic neurocircuitry, thereby contributing to imbalances in appetite and energy homeostasis and increasing the risk of obesity in subsequent generations. Early in postnatal life, the neuronal projections responsible for energy homeostasis develop in response to appetite-related peptides such as leptin. To date, no study characterises the genome-wide transcriptional changes that occur in response to exposure to high fat diet during this critical window. We explored the effects of maternal high fat diet consumption on hypothalamic gene expression in Sprague Dawley rat offspring at postnatal day 10. RNA-sequencing enabled discovery of differentially expressed genes between offspring of dams fed a high fat diet and offspring of control diet fed dams. Female high fat diet offspring displayed altered expression of 86 genes (adjusted P-value<0.05), including genes coding for proteins of the extra cellular matrix, particularly Collagen 1a1 (Col1a1), Col1a2, Col3a1, and the imprinted Insulin-like growth factor 2 (Igf2) gene. Male high fat diet offspring showed significant changes in collagen genes (Col1a1 and Col3a1) and significant upregulation of two genes involved in regulation of dopamine availability in the brain, tyrosine hydroxylase (Th) and dopamine reuptake transporter Slc6a3 (also known as Dat1). Transcriptional changes were accompanied by increased body weight, body fat and body length in the high fat diet offspring, as well as altered blood glucose and plasma leptin. Transcriptional changes identified in the hypothalamus of offspring of high fat diet mothers could alter neuronal projection formation during early development leading to abnormalities in the neuronal circuitry controlling appetite in later life, hence priming offspring to the development of obesity.

Colonization with Escherichia coli EC 25 protects neonatal rats from necrotizing enterocolitis.

Necrotizing enterocolitis (NEC) is a significant cause of morbidity and mortality in premature infants; yet its pathogenesis remains poorly understood. To evaluate the role of intestinal bacteria in protection against NEC, we assessed the ability of naturally occurring intestinal colonizer E. coli EC25 to influence composition of intestinal microbiota and NEC pathology in the neonatal rat model. Experimental NEC was induced in neonatal rats by formula feeding/hypoxia, and graded histologically. Bacterial populations were characterized by plating on blood agar, scoring colony classes, and identifying each class by sequencing 16S rDNA. Binding of bacteria to, and induction of apoptosis in IEC-6 enterocytes were examined by plating on blood agar and fluorescent staining for fragmented DNA. E. coli EC 25, which was originally isolated from healthy rats, efficiently colonized the intestine and protected from NEC following introduction to newborn rats with formula at 106 or 108 cfu. Protection did not depend significantly on EC25 inoculum size or load in the intestine, but positively correlated with the fraction of EC25 in the microbiome. Introduction of EC25 did not prevent colonization with other bacteria and did not significantly alter bacterial diversity. EC25 neither induced cultured enterocyte apoptosis, nor protected from apoptosis induced by an enteropathogenic strain of Cronobacter muytjensii. Our results show that E. coli EC25 is a commensal strain that efficiently colonizes the neonatal intestine and protects from NEC.

Topical anaesthesia reduces sensitivity of castration wounds in neonatal piglets.

The aim of this study was to do determine the efficacy of the topical anaesthetic Tri-Solfen® in the amelioration of the pain of castration in piglets. The trial was conducted over a three day period, and blocked across six litters with 12 piglets treated on days one and two, and 16 on day three. The piglets were randomly allocated by weight and litter to 1 of 4 treatment groups: (i) sham castration (SHAM; n = 10); (ii) surgical castration with no anaesthetic intervention (CAST; n = 10); (iii) surgical castration with post-operative topical anaesthesia (TRI; n = 10); (iv) surgical castration with a pre-operative intra-testicular lignocaine hydrochloride injection (LIG; n = 10). Wound sensitivity testing involved von Frey monofilaments of weights 4g and 300g, and an 18 gauge needle, used to stimulate the wound and surrounding skin respectively, at various pre-determined sites. Neonatal piglets receiving topical anaesthesia (Tri-Solfen®) spray into castration wounds had significantly lower wound sensitivity responses for up to 4h, compared to those castrated following intra-testicular lignocaine injection or those with no treatment. The use of topical anaesthetic is suggested as a practical and affordable method of improving piglet welfare during castration.

Infanticide in chimpanzees: Taphonomic case studies from Gombe.

We present a study of skeletal damage to four chimpanzee (Pan troglodytes) infanticide victims from Gombe National Park, Tanzania. Skeletal analysis may provide insight into the adaptive significance of infanticide by examining whether nutritional benefits sufficiently explain infanticidal behavior. The nutritional hypothesis would be supported if bone survivorship rates and skeletal damage patterns are comparable to those of monkey prey. If not, other explanations, such as the resource competition hypothesis, should be considered.

Exposure to intrauterine inflammation alters metabolomic profiles in the amniotic fluid, fetal and neonatal brain in the mouse.

Exposure to prenatal inflammation is associated with diverse adverse neurobehavioral outcomes in exposed offspring. The mechanism by which inflammation negatively impacts the developing brain is poorly understood. Metabolomic profiling provides an opportunity to identify specific metabolites, and novel pathways, which may reveal mechanisms by which exposure to intrauterine inflammation promotes fetal and neonatal brain injury. Therefore, we investigated whether exposure to intrauterine inflammation altered the metabolome of the amniotic fluid, fetal and neonatal brain. Additionally, we explored whether changes in the metabolomic profile from exposure to prenatal inflammation occurs in a sex-specific manner in the neonatal brain.

Triglyceride concentrations in neonatal foals: Serial measurement and effects of age and illness.

Few studies have evaluated the effects of age and illness on serum triglyceride concentrations in neonatal foals. The objectives of this study were to evaluate triglyceride concentrations in neonatal foals and their dams through serial measurement immediately postpartum and at 1-2 days and 10-12 days of age, as well as to measure them in sick foals. Serially measured serum triglycerides in seven healthy foals varied with age. Median (range) triglyceride concentrations were 28mg/dL (12-50mg/dL), 89mg/dL (51-264mg/dL), and 60mg/dL (28-135mg/dL) immediately postpartum, at 1-2 days of age, and 10-12 days of age, respectively (P<0.001). Triglyceride concentrations varied hourly by up to 117mg/dL in individual foals. The dams had lower triglycerides (median, 20mg/dL; range, 12-49mg/dL) than the foals, once foals were >24h old. Sick foals <24h old had lower triglycerides than sick foals aged 1-7 days (median, 41mg/dL [range, 16-116]; median, 110mg/dL [range, 24-379mg/dL]; P<0.001). Age and triglyceride concentration showed a non-linear association independent of foal health status (P=0.01). Sick foals with positive bacterial cultures had higher triglycerides than those with negative cultures (median, 111mg/dL [range, 10-379mg/dL] and median 53mg/dL [range, 17-271mg/dL], respectively; P=0.033). Nonsurvivors had higher triglycerides than survivors (median, 116mg/dL [range, 41-379mg/dL] and median, 55mg/dL [range, 10-311mg/dL], respectively; P=0.04). In conclusion, triglycerides were highest in healthy neonatal foals aged 1-2 days, and in nonsurviving sick foals and those with positive bacterial cultures. Age was associated with triglyceride concentration regardless of health status.

A newborn infant chimpanzee snatched and cannibalized immediately after birth: Implications for "maternity leave" in wild chimpanzee.

This study reports on the first observed case of a wild chimpanzee infant being snatched immediately after delivery and consequently cannibalized by an adult male in the Mahale Mountains, Tanzania. We demonstrate "maternity leave" from long-term data from the Mahale M group and suggest that it functions as a possible counterstrategy of mother chimpanzees against the risk of infanticide soon after delivery.

Role of mycotoxins in herds with and without problems with tail necrosis in neonatal pigs.

This study aimed to investigate a possible involvement of mycotoxins in neonatal tail necrosis in piglets. Ten affected and 10 non-affected farms were selected. Sow feed samples were analysed for the presence of 23 mycotoxins by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Blood plasma samples of sows and their piglets were analysed for the presence of deoxynivalenol (DON), de-epoxydeoxynivalenol, T-2 and HT-2 toxin, zearalenone, alfa-zearalenol, and beta-zearalenol, using LC-MS/MS. Additionally, high-resolution mass spectrometry (HRMS) was performed to detect DON-glucuronide (DON-Glca). There was a significant difference between case herds and control herds for mean DON concentrations in feed and sow plasma. For piglet samples, concentrations of DON were above the limit of quantification of 0.1 ng/ml in only 12 samples. Positive correlations were found between DON concentrations in sow feed and plasma of sows; DON concentration in sow feed and DON-Glca concentration in plasma of sows; and between DON and DON-Glca concentration in sow-plasma. In conclusion, high prevalence of DON in feed samples was found, with significantly higher concentrations in case herds, as well as the presence of DON and DON-Glca in sow plasma. Additional research is needed to identify risk factors, including within-herd factors, associated with neonatal tail necrosis in piglets.

Protective effect of cyanidin-3-O-glucoside on neonatal porcine islets.

Oxidative stress is a major cause of islet injury and dysfunction during isolation and transplantation procedures. Cyanidin-3-O-glucoside (C3G), which is present in various fruits and vegetables especially in Chinese bayberry, shows a potent antioxidant property. In this study, we determined whether C3G could protect neonatal porcine islets (NPI) from reactive oxygen species (H2O2)-induced injury in vitro and promote the function of NPI in diabetic mice. We found that C3G had no deleterious effect on NPI and that C3G protected NPI from damage induced by H2O2 Significantly higher hemeoxygenase-1 (HO1) gene expression was detected in C3G-treated NPI compared to untreated islets before and after transplantation (P < 0.05). Western blot analysis showed a significant increase in the levels of phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2) and phosphatidylinositol 3-kinase (PI3K/Akt) proteins in C3G-treated NPI compared to untreated islets. C3G induced the nuclear translocation of nuclear erythroid 2-related factor 2 (NRF2) and the significant elevation of HO1 protein. Recipients of C3G-treated NPI with or without C3G-supplemented drinking water achieved normoglycemia earlier compared to recipients of untreated islets. Mice that received C3G-treated islets with or without C3G-supplemented water displayed significantly lower blood glucose levels at 5-10 weeks post-transplantation compared to mice that received untreated islets. Mice that received C3G-treated NPI and C3G-supplemented drinking water had significantly (P < 0.05) lower blood glucose levels at 7 and 8 weeks post-transplantation compared to mice that received C3G-treated islets. These findings suggest that C3G has a beneficial effect on NPI through the activation of ERK1/2- and PI3K/AKT-induced NRF2-mediated HO1 signaling pathway.

Female reproductive success and calf survival in a North Sea coastal bottlenose dolphin (Tursiops truncatus) population.

Between-female variation in reproductive output provides a strong measure of individual fitness and a quantifiable measure of the health of a population which may be highly informative to management. In the present study, we examined reproductive traits in female bottlenose dolphins from the east coast of Scotland using longitudinal sightings data collected over twenty years. From a total of 102 females identified between 1997 and 2016, 74 mothers produced a collective total of 193 calves. Females gave birth from 6 to 13 years of age with a mean age of 8. Calves were produced during all study months, May to October inclusive, but showed a seasonal birth pulse corresponding to the regional peak in summer water temperatures. Approximately 83% (n = 116) of the calves of established fate were successfully raised to year 2-3. Of the known mortalities, ~45% were first-born calves. Calf survival rates were also lower in multiparous females who had previously lost calves. A mean inter-birth interval (IBI) of 3.80 years (n = 110) and mean fecundity of 0.16 was estimated for the population. Calf loss resulted in shortened IBIs, whilst longer IBIs were observed in females assumed to be approaching reproductive senescence. Maternal age and size, breeding experience, dominance, individual associations, group size and other social factors, were all concluded to influence reproductive success (RS) in this population. Some females are likely more important than others for the future viability of the population. Consequently, a better knowledge of the demographic groups containing those females showing higher reproductive success would be highly desirable for conservation efforts aimed at their protection.

Relative tooth size at birth in primates: Life history correlates.

Dental eruption schedules have been closely linked to life history variables. Here we examine a sample of 50 perinatal primates (28 species) to determine whether life history traits correlate with relative tooth size at birth.

Bone up: craniomandibular development and hard-tissue biomineralization in neonate mice.

The presence of regional variation in the osteogenic abilities of cranial bones underscores the fact that the mechanobiology of the mammalian skull is more complex than previously recognized. However, the relationship between patterns of cranial bone formation and biomineralization remains incompletely understood. In four strains of mice, micro-computed tomography was used to measure tissue mineral density during perinatal development in three skull regions (calvarium, basicranium, mandible) noted for variation in loading environment, embryological origin, and ossification mode. Biomineralization levels increased during perinatal ontogeny in the mandible and calvarium, but did not increase in the basicranium. Tissue mineral density levels also varied intracranially, with density in the mandible being highest, in the basicranium intermediate, and in the calvarium lowest. Perinatal increases in, and elevated levels of, mandibular biomineralization appear related to the impending postweaning need to resist elevated masticatory stresses. Similarly, perinatal increases in calvarial biomineralization may be linked to ongoing brain expansion, which is known to stimulate sutural bone formation in this region. The lack of perinatal increase in basicranial biomineralization could be a result of earlier developmental maturity in the cranial base relative to other skull regions due to its role in supporting the brain's mass throughout ontogeny. These results suggest that biomineralization levels and age-related trajectories throughout the skull are influenced by the functional environment and ontogenetic processes affecting each region, e.g., onset of masticatory loads in the mandible, whereas variation in embryology and ossification mode may only have secondary effects on patterns of biomineralization. Knowledge of perinatal variation in tissue mineral density, and of normal cranial bone formation early in development, may benefit clinical therapies aiming to correct developmental defects and traumatic injuries in the skull, and more generally characterize loading environments and skeletal adaptations in mammals by highlighting the need for multi-level analyses for evaluating functional performance of cranial bone.

Retinal VEGF levels correlate with ocular circulation measured by a laser speckle-micro system in an oxygen-induced retinopathy rat model.

We used a Laser speckle flowgraphy (LSFG)-micro system to examine the relationship between ocular blood flow and retinal vascular endothelial growth factor (VEGF) at retinopathy onset in oxygen-induced ischemic retinopathy (OIR) model rats.

Prevalence and calf-level risk factors for failure of passive transfer in dairy calves in New Zealand.

AIM To determine the prevalence of failure of passive transfer (FPT) in dairy calves in New Zealand and to assess a range of potential calf-level risk factors for their association with FPT. METHOD Between July and September 2015, 107 dairy farms in nine regions across New Zealand were visited on three occasions: before 20% of cows had calved, when ∼50% of cows had calved, and when the last 10-20% of cows were calving. Farms were randomly selected from the client list of participating veterinary practices. On each farm, blood samples were collected from 10-20 calves between 24 hours and 8 days old and concentrations of total protein (TP) in serum determined. Calf identification, date of birth, dam's age and breed, details of colostrum feeding and the farmer's health assessment of the calf on arrival at the calf rearing facility were recorded. Concentrations of TP in serum ≤52 g/L were used to define FPT. RESULTS Of 3,819 calves sampled, 1,263 (33 (95% CI=31.6-34.6)%) were diagnosed with FPT, and the prevalence within farms ranged from 5-83%. Calf-level risk factors that were associated with increased odds of FPT were increasing age of dam (OR=1.12; 95% CI=1.08-1.16), calves recorded as unhealthy compared to healthy upon arrival at the calf rearing facility (OR=2.59; 95% CI=1.60-4.19), calves sampled on farms in Otago (OR=2.85; 95% CI=1.42-5.74) and Southland (OR 2.31; 95% CI=1.15-4.67) compared to the Far North, and calves sampled in the middle compared to the early calving period (OR=1.24; 95% CI=1.03-1.51). CONCLUSION On the farms visited FPT was diagnosed in one third of dairy calves. Calf-level risk factors for FPT included age of dam, health status of calf, region, and time of the calving period. Further investigations are required to establish why these are associated with increased odds of FPT. CLINICAL RELEVANCE These results provide a baseline for benchmarking, which, alongside the reported calf-level risk factors, can be used by veterinarians to improve colostrum management.

Effects of feeding pregnant beef cows selenium-enriched alfalfa hay on selenium status and antibody titers in their newborn calves.

In newborn dairy calves, it has been demonstrated that supranutritional maternal and colostral Se supplementation using Se yeast or sodium selenite, respectively, improves passive transfer of IgG. In beef cattle, agronomic biofortification with Se is a more practical alternative for Se supplementation, whereby the Se concentration of hay is increased through the use of Se-containing fertilizer amendments. It has been previously demonstrated that agronomic Se biofortification is an effective strategy to improve immunity and performance in Se-replete weaned beef calves. The objective of this experiment was to determine the effects of feeding beef cows Se-enriched alfalfa () hay during the last 8 to 12 wk of gestation on passive transfer of antibodies to calves. At 10 wk ± 16 d before calving, 45 cows were assigned to 1 of 3 treatment groups with 3 pens (5 cows/pen) per treatment: Control cows were fed non-Se-fortified alfalfa hay plus a mineral supplement containing 120 mg/kg Se from sodium selenite, Med-Se cows were fed alfalfa hay fertilized with 45.0 g Se/ha as sodium selenate, and High-Se cows were fed alfalfa hay fertilized with 89.9 g Se/ha as sodium selenate; both the Med-Se and the High-Se groups received mineral supplement without added Se. Colostrum and whole blood (WB) were collected from cows at calving, and WB was collected from calves within 2 h of calving and at 12, 24, 36, and 48 h of age. Concentrations of IgG1 and J-5 antibody in cow colostrum and calf serum were quantified using ELISA procedures. Selenium concentrations linearly increased in WB ( < 0.001) and colostrum ( < 0.001) of cows and in WB of newborn calves ( < 0.001) with increasing Se concentration in alfalfa hay. Colostrum concentrations of IgG1 ( = 0.03) were increased in cows fed Se-biofortified alfalfa hay, but J-5 antibody ( = 0.43) concentrations were not. Calf serum IgG1 ( = 0.43) and J-5 antibody ( = 0.44) concentrations during the first 48 h of age were not affected by prior Se treatment of cows. These data suggest that feeding Se-biofortified alfalfa hay promotes the accumulation of Se and antibodies in colostrum but does not affect short-term serum antibody concentrations in calves.

Steroids, steroid precursors, and neuroactive steroids in critically ill equine neonates.

Hypothalamic-pituitary-adrenal axis (HPAA) dysfunction has been associated with sepsis and mortality in foals. Most studies have focused on cortisol, while other steroids have not been investigated. The objectives of this study were to characterise the adrenal steroid and steroid precursor response to disease and to determine their association with the HPAA response to illness, disease severity, and mortality in hospitalised foals. All foals (n=326) were classified by two scoring systems into three categories: based on the sepsis score (septic, sick non-septic [SNS] and healthy) and the foal survival score (Group 1: 3-18%; Group 2: 38-62%; Group 3: 82-97% likelihood of survival). Blood concentrations of adrenocorticotropic hormone (ACTH) and steroids were determined by immunoassays. ACTH-cortisol imbalance (ACI) was defined as a high ACTH/cortisol ratio. Septic foals had higher ACTH, cortisol, progesterone, 17α-OH-progesterone, pregnenolone, and androstenedione concentrations as well as higher ACTH/cortisol, ACTH/progesterone, ACTH/aldosterone, and ACTH/DHEAS ratios than SNS and healthy foals (P<0.01). Foals with DHEAS of 0.4-5.4ng/mL were more likely to have ACI (OR=2.5). Foals in Group 1 had higher ACTH, aldosterone, progesterone, and cortisol concentrations as well as ACTH/cortisol, ACTH/progesterone, and ACTH/DHEAS ratios than foals in Groups 2 and 3 (P<0.01). High progesterone concentrations were associated with non-survival and the cutoff value below which survival could be predicted was 23.5ng/mL, with 75% sensitivity and 72% specificity. In addition to cortisol, the response to the stress of illness in foals is characterised by the release of multiple adrenal steroids. DHEAS and progesterone were good predictors of HPAA dysfunction and outcome in hospitalised foals.

Supplementation with Lactobacillus paracasei or Pediococcus pentosaceus does not prevent diarrhoea in neonatal pigs infected with Escherichia coli F18.

Infectious diarrhoea is a worldwide problem in newborns. Optimal bacterial colonisation may enhance gut maturation and protect against pathogenic bacteria after birth. We hypothesised that lactic acid bacteria (LAB) administration prevents pathogen-induced diarrhoea in formula-fed newborns. Newborn caesarean-delivered, colostrum-deprived term piglets on parenteral nutrition for the first 15 h, were used as models for sensitive newborn infants. A commercially available probiotic strain, Lactobacillus paracasei F19 (LAP, 2·6×108 colony-forming units (CFU)/kg per d) and a novel LAB isolate, Pediococcus pentosaceus (PEP, 1·3×1010 CFU/kg per d), were administered for 5 d with or without inoculation of the porcine pathogen, Escherichia coli F18 (F18, 1010 CFU/d). This resulted in six treatment groups: Controls (n 9), LAP (n 10), PEP (n 10), F18 (n 10), F18-LAP (n 10) and F18-PEP (n 10). The pathogen challenge increased diarrhoea and density of F18 in the intestinal mucosa (P<0·05). LAB supplementation further increased the diarrhoea score, relative to F18 alone (P<0·01). Intestinal structure and permeability were similar among groups, whereas brush border enzymes were affected in variable intestinal regions with decreased activities in most cases after F18 and LAB inoculation. Bacterial density in colon mucosa increased after F18 inoculation (P<0·05) but was unaffected by LAB supplementation. In colon contents, acetic and butyric acids were increased by PEP (P<0·05). The LAB used in this study failed to reduce E. coli-induced diarrhoea in sensitive newborn pigs. In vulnerable newborns there may be a delicate balance among bacterial composition and load, diet and the host. Caution may be required when administering LAB to compromised newborns suffering from enteric infections.

Diagnosis and Treatment Considerations for Nonphyseal Long Bone Fractures in the Foal.

Many long bone fractures that are not considered repairable in the adult horse are repairable in the foal. This is largely because of reduced patient size and more rapid healing in the foal. When there is no articular communication, the long-term prognosis for athletic function can be very good. Emergency care and transport of the foal with a long bone fracture is different than the adult.

Physeal Fractures in Foals.

Physeal fractures are common musculoskeletal injuries in foals and should be included as a differential diagnosis for the lame or nonweightbearing foal. Careful evaluation of the patient, including precise radiographic assessment, is paramount in determining the options for treatment. Prognosis mostly depends on the patient's age, weight, and fracture location and configuration.

Routine Orthopedic Evaluation in Foals.

In order to recognize abnormalities on the physical evaluation, it is mandatory to understand normal developmental variations of the musculoskeletal system. Many abnormalities are self-limiting and, therefore, it is important to recognize which problems require intervention for a successful outcome and which may be complicated by treatment. The importance of a complete and thorough physical evaluation cannot be overemphasized and is the most productive diagnostic tool for recognizing most abnormalities of the skeletal system whether as a component of an after-foaling examination or for lameness or conformation evaluation in foals of all ages.

Ultrasonographic assessment of the atlanto-occipital space in healthy Thoroughbred foals and Thoroughbred foals with neonatal maladjustment syndrome.

Ultrasonography of the atlanto-occipital (AO) space may be useful as a non-invasive diagnostic tool in neonatal foals. The aims of the study were establish a range of values for ultrasonographic measurements of the AO space in healthy Thoroughbred foals and to compare these variables in healthy foals with foals diagnosed with neonatal maladjustment syndrome (NMS). Ultrasonography of the AO space was performed on 38 healthy Thoroughbred foals and 28 Thoroughbred foals with NMS≤4days of age. Transverse image spinal cord height (P=0.001), width (P<0.001) and spinal cord cross sectional area (P<0.001), and longitudinal image dorsoventral diameter of the ventral spinal artery, were significantly smaller in foals with NMS than in healthy foals. Ratios of spinal canal to cord width and cross sectional area were significantly smaller in healthy foals than in foals with NMS (P<0.001). Spinal canal variables were not significantly different between groups. Several ultrasonographic measurements of the AO space were significantly different between healthy foals and foals with NMS. Further investigation is warranted to investigate the clinical application of this technique.

Effects of iron supplementation on growth, gut microbiota, metabolomics and cognitive development of rat pups.

Iron deficiency is common during infancy and therefore iron supplementation is recommended. Recent reports suggest that iron supplementation in already iron replete infants may adversely affect growth, cognitive development, and morbidity.

Importance of neonatal immunoglobulin transfer for hippocampal development and behaviour in the newborn pig.

Neurological disorders are among the main clinical problems affecting preterm children and often result in the development of communication and learning disabilities later in life. Several factors are of importance for brain development, however the role of immunoglobulins (passive immunity transfer) has not yet been investigated. Piglets are born agammaglobulinemic, as a result of the lack of transfer of maternal immunoglobulins in utero, thus, they serve as an ideal model to mimic the condition of immunoglobulin deficiency in preterm infants. Thirty six, unsuckled newborn piglets were fed an infant formula or colostrum and supplemented orally or intravenously with either species-specific or foreign immunoglobulin and then compared to both newborn and sow-reared piglets. Two days after the piglets were born behavioural tests (novel recognition and olfactory discrimination of conspecifics scent) were performed, after which the piglets were sacrificed and blood, cerebrospinal fluid and hippocampi samples were collected for analyses. Both parameters of neuronal plasticity (neuronal maturation and synapse-associated proteins) and behavioural test parameters appeared to be improved by the appearance of species-specific porcine immunoglulin in the circulation and cerebrospinal fluid of the piglets. In conclusion, we postulate possible positive clinical effects following intravenous infusion of human immunoglobulin in terms of neuronal plasticity and cognitive function in preterm infants born with low blood immunoglobulin levels.

Dicephalic Parapagus Conjoined Twins in a Large Fruit-eating Bat, Genus Artibeus (Chiroptera, Phyllostomidae).

Conjoined twinning is an embryological anomaly rarely reported in wild mammals and with only two previous records in Chiroptera. Here, we report a case of dicephalic parapagus conjoined twins in the Neotropical phyllostomid genus Artibeus. These twins are males and present separated heads and necks, but a conjoined trunk with an expanded upper thoracic region. They developed two complete forelimbs and two complete hindlimbs, all laterally to the trunk. There is a volume in the upper midback and between the heads that resembles a third rudimentary medial forelimb, but X-ray images only suggest the presence of medial skeletal elements of the pectoral girdle (clavicle and scapulae) in this region. The X-ray images also show that vertebral columns run separated from head until the base of lumbar region, where they form a single structure. Using ultrasound images, we detected the presence of two similarly sized and apparently separated hearts. The accumulation of study cases like this will help in the understanding of patterns and process behind this phenomena, and collection material plays a key role in this context.

Unusually high-pitched neonate distress calls of the open-habitat Mongolian gazelle (Procapra gutturosa) and their anatomical and hormonal predictors.

In neonate ruminants, the acoustic structure of vocalizations may depend on sex, vocal anatomy, hormonal profiles and body mass and on environmental factors. In neonate wild-living Mongolian gazelles Procapra gutturosa, hand-captured during biomedical monitoring in the Daurian steppes at the Russian-Mongolian border, we spectrographically analysed distress calls and measured body mass of 22 individuals (6 males, 16 females). For 20 (5 male, 15 female) of these individuals, serum testosterone levels were also analysed. In addition, we measured relevant dimensions of the vocal apparatus (larynx, vocal folds, vocal tract) in one stillborn male Mongolian gazelle specimen. Neonate distress calls of either sex were high in maximum fundamental frequency (800-900 Hz), but the beginning and minimum fundamental frequencies were significantly lower in males than in females. Body mass was larger in males than in females. The levels of serum testosterone were marginally higher in males. No correlations were found between either body mass or serum testosterone values and any acoustic variable for males and females analysed together or separately. We discuss that the high-frequency calls of neonate Mongolian gazelles are more typical for closed-habitat neonate ruminants, whereas other open-habitat neonate ruminants (goitred gazelle Gazella subgutturosa, saiga antelope Saiga tatarica and reindeer Rangifer tarandus) produce low-frequency (<200 Hz) distress calls. Proximate cause for the high fundamental frequency of distress calls of neonate Mongolian gazelles is their very short, atypical vocal folds (4 mm) compared to the 7-mm vocal folds of neonate goitred gazelles, producing distress calls as low as 120 Hz.

Natural killer T cell sensitization during neonatal respiratory syncytial virus infection induces eosinophilic lung disease in re-infected adult mice.

Respiratory syncytial virus (RSV) is a major viral pathogen that causes severe lower respiratory tract infections in infants and the elderly worldwide. Infants with severe RSV bronchiolitis tend to experience more wheezing and asthma in later childhood. Because invariant natural killer T (iNKT) cells are associated with the asthma pathology, we investigated whether neonatal iNKT cells are involved in the aggravation of pulmonary diseases following RSV infection in mice. Intranasal exposure to the iNKT cell ligand α-galactosylceramide (α-GC) with RSV primary infection in neonatal mice elicited neither cytokine production (except for a slight increase of IL-5) nor pulmonary eosinophilia, despite the presence of both CD1d+ cells and NKT cells. Interestingly, in adult mice re-infected with RSV, neonatal iNKT cell sensitization by α-GC during RSV primary infection resulted in much higher levels of pulmonary Th2 cytokines and elevated eosinophilia with airway hyperresponsiveness, whereas this was not observed in cd1d knockout mice. In contrast, α-GC priming of adults during RSV re-infection did not induce more severe airway symptoms than RSV re-infection in the absence of α-GC. α-GC co-administration during RSV primary infection facilitated RSV clearance regardless of age, but viral clearance following re-infection was not iNKT cell-dependent. This study clearly demonstrates that RSV-induced immune responses can be altered by iNKT cells, suggesting that neonatal iNKT cell sensitization during RSV primary infection is associated with exacerbation of pulmonary diseases following RSV re-infection in adulthood.

Biomedical research: Premature lambs grown in a bag.

Intraperitoneal Administration of Ethanol as a Means of Euthanasia for Neonatal Mice (Mus musculus).

The humane euthanasia of animals in research is of paramount importance. Neonatal mice frequently respond differently to euthanasia agents when compared with adults. The AVMA's Guidelines for the Euthanasia of Animals includes intraperitoneal injection of ethanol as "acceptable with conditions," and recent work confirmed that this method is appropriate for euthanizing adult mice, but neonatal mice have not been tested. To explore this method in neonatal mice, mouse pups (C57BL/6 and CD1, 162 total) were injected with 100% ethanol, a pentobarbital-phenytoin combination, or saline at 7, 14, 21, 28, or 35 d of age. Electrocardiograms, respiratory rates, and times to loss of righting reflex and death were recorded. Time to death (TTD) differed significantly between ethanol and pentobarbital-phenytoin at 7, 14, and 21 d and between ethanol groups at 7, 14, and 21 d compared with 35 d. The average TTD (± 1 SD) for ethanol-injected mice were: 7 d, 70.3 ± 39.8 min; 14 d, 51.7 ± 30.5 min; 21 d, 32.3 ± 20.8 min, 28 d, 14.0 ± 15.2; and 35 d, 4.9 ± 1.4. Mean TTD in pentobarbital-phenytoin-injected mice were: 7 d, 2.8 ± 0.4 min; 14 d, 2.9 ± 0.5 min; 21 d, 3.9 ± 1.2 min; 28 d, 3.9 ± 0.7 min; and 35 d, 4.4 ± 0.5. Although TTD did not differ between ethanol and pentobarbital-phenytoin at 28 d of age, the TTD in 3 of 12 mice was longer than 15 min after ethanol administration at this age. Therefore, ethanol should not be used as a method of euthanasia for mice younger than 35 d, because the criteria for humane euthanasia were met only in mice 35 d or older.

Spatial regulation of bone morphogenetic proteins (BMPs) in postnatal articular and growth plate cartilage.

Articular and growth plate cartilage both arise from condensations of mesenchymal cells, but ultimately develop important histological and functional differences. Each is composed of three layers-the superficial, mid and deep zones of articular cartilage and the resting, proliferative and hypertrophic zones of growth plate cartilage. The bone morphogenetic protein (BMP) system plays an important role in cartilage development. A gradient in expression of BMP-related genes has been observed across growth plate cartilage, likely playing a role in zonal differentiation. To investigate the presence of a similar expression gradient in articular cartilage, we used laser capture microdissection (LCM) to separate murine growth plate and articular cartilage from the proximal tibia into their six constituent zones, and used a solution hybridization assay with color-coded probes (nCounter) to quantify mRNAs for 30 different BMP-related genes in each zone. In situ hybridization and immunohistochemistry were then used to confirm spatial expression patterns. Expression gradients for Bmp2 and 6 were observed across growth plate cartilage with highest expression in hypertrophic zone. However, intracellular BMP signaling, assessed by phospho-Smad1/5/8 immunohistochemical staining, appeared to be higher in the proliferative zone and prehypertrophic area than in hypertrophic zone, possibly due to high expression of Smad7, an inhibitory Smad, in the hypertrophic zone. We also found BMP expression gradients across the articular cartilage with BMP agonists primarily expressed in the superficial zone and BMP functional antagonists primarily expressed in the deep zone. Phospho-Smad1/5/8 immunohistochemical staining showed a similar gradient. In combination with previous evidence that BMPs regulate chondrocyte proliferation and differentiation, the current findings suggest that BMP signaling gradients exist across both growth plate and articular cartilage and that these gradients may contribute to the spatial differentiation of chondrocytes in the postnatal endochondral skeleton.

Hypothalamic effects of neonatal diet: reversible and only partially leptin dependent.

Early life diet influences metabolic programming, increasing the risk for long-lasting metabolic ill health. Neonatally overfed rats have an early increase in leptin that is maintained long term and is associated with a corresponding elevation in body weight. However, the immediate and long-term effects of neonatal overfeeding on hypothalamic anorexigenic pro-opiomelanocortin (POMC) and orexigenic agouti-related peptide (AgRP)/neuropeptide Y (NPY) circuitry, and if these are directly mediated by leptin, have not yet been examined. Here, we examined the effects of neonatal overfeeding on leptin-mediated development of hypothalamic POMC and AgRP/NPY neurons and whether these effects can be normalised by neonatal leptin antagonism in male Wistar rats. Neonatal overfeeding led to an acute (neonatal) resistance of hypothalamic neurons to exogenous leptin, but this leptin resistance was resolved by adulthood. While there were no effects of neonatal overfeeding on POMC immunoreactivity in neonates or adults, the neonatal overfeeding-induced early increase in arcuate nucleus (ARC) AgRP/NPY fibres was reversed by adulthood so that neonatally overfed adults had reduced NPY immunoreactivity in the ARC compared with controls, with no further differences in AgRP immunoreactivity. Short-term neonatal leptin antagonism did not reverse the excess body weight or hyperleptinaemia in the neonatally overfed, suggesting factors other than leptin may also contribute to the phenotype. Our findings show that changes in the availability of leptin during early life period influence the development of hypothalamic connectivity short term, but this is partly resolved by adulthood indicating an adaptation to the metabolic mal-programming effects of neonatal overfeeding.