The fascinating microbes and their impact on neonatal dogs and cats - A review.

Recent literature data indicate that canine and feline neonates are not born in a sterile environment as it was stated previously. The acquisition, colonisation and maintenance of the early life microbiota of healthy fetuses is a rapidly developing research area. In humans, the natural healthy infant microbiome plays an essential role in health and its assembly is determined by the maternal-offspring exchanges of microbes. Even though this topic is becoming more and more important in dogs and cats, the exact role of the neonatal microbiome is not yet fully known in animals. This review summarises the current knowledge of the normal physiological neonatal microbiome in healthy puppies and kittens.

Staphylococcus pseudintermedius septicemia in puppies after elective cesarean section: confirmed transmission via dam's milk.

BACKGROUND: In humans, transmission of bacteria causing fatal sepsis in the neonates through mother's milk has been reported. In dogs, it is believed that bacteria from canine milk are not the primary cause of neonatal infections. Staphylococcus pseudintermedius is colonizing the skin and mucocutaneous junctions in adult dogs and can act as an opportunistic pathogen. This bacterium was previously isolated from the canine milk and, although, its transmission from the dam's milk to the newborn puppies causing a neonatal sepsis was suggested, this hypothesis has not been confirmed. CASE PRESENTATION: A 4.5-year-old healthy Boston terrier dam had an elective cesarean section, delivering five normal puppies and one dead runt. Next day, two puppies developed pustules on their legs and around the muzzle. After two more days, strings of blood were noticed in the stool of the biggest puppy that suddenly died later that night. The same day, blood became visible in the feces of all other puppies. Necropsy of the dead puppy revealed a distended abdomen, catarrhal gastroenteritis with lymphadenopathy, dark red and slightly firm lung, mild dilatation of the right heart chamber and congestion of the liver, spleen, pancreas and meninges. The thoracic cavity contained white-yellow slightly opaque exudate, and there was transudate in the abdominal cavity. Histopathology revealed an acute interstitial pneumonia and multifocal myocardial necrosis with mineralization. Bacteriology of the internal organs, body cavity effusions of the dead puppy and dam's milk revealed a diffuse growth of S. pseudintermedius in pure culture. Whole genome sequencing (WGS) revealed that all isolates belonged to the sequence type 241 and differed in 2-5 single nucleotide polymorphisms; thus, the epidemiological link between the outbreak-associated isolates was confirmed. CONCLUSIONS: This is the first report of a confirmed transmission of S. pseudintermedius through dam's milk causing a neonatal sepsis in a puppy after an elective cesarean section. The epidemiological link between S. pseudintermedius isolates obtained from dam's milk and internal organs of the affected puppy was confirmed by WGS. Our findings indicate that milk of healthy dams can serve as a reservoir of bacteria that can cause fatal sepsis in the newborn puppies.

Identification of the Identical Human Mutation in ACVR1 in 2 Cats With Fibrodysplasia Ossificans Progressiva.

Two domestic shorthair cats, 1 intact female and 1 intact male, presented with progressive limb lameness and digital deformities at 4 and 6 months of age. Stiffness and swelling of the distal thoracic and pelvic limb joints progressed to involve hip and shoulder joints, resulting in reduced mobility. Radiographs in both cats and computed tomography of the male cat revealed ankylosing, polyarticular deposits of extracortical heterotopic bone spanning multiple axial and appendicular joints, extending into adjacent musculotendinous tissues. All findings supported fibrodysplasia ossificans progressiva (FOP), a disorder characterized by toe malformations and progressive heterotopic ossification in humans. In both cats, molecular analyses revealed the same heterozygous mutation in the activin A receptor type I (ACVR1) gene that occurs in humans with FOP. Several reports of heterotopic ossification in cats exist, but this is the first one to identify clinical FOP in 2 cats with the identical mutation that occurs in >95% of humans with FOP.

Beyond Birth: Pioneering Insights into Colostrum Quality Variation among Bitches with Different Types of Parturition.

This study deals with the immunological quality of canine colostrum in relation to the different parturition modes in bitches. It included four groups of bitches, who underwent vaginal parturition (VP), emergency cesarean section (EM-CS), elective cesarean section (EL-CS), or elective cesarean section with aglepristone injection 24 h prior to surgery (EL-A). Colostrum samples from 40 bitches of 18 breeds were taken immediately after the birth of the first puppy or directly after surgery, and four hours later. The concentrations of immunoglobulin G (IgG) and IgA were measured using ELISA tests. The initial IgG concentration was 18.3 ± 10.2 g/L, and the IgA concentration was 13.7 ± 5.8 g/L, respectively. Significantly lower IgG and IgA concentrations were observed in the EL-CS group compared to other groups. The administration of aglepristone led to an increase in IgG and IgA concentrations. Despite the lower immunoglobulin levels with EL-CS, the survival rate of the puppies was not affected. The study shows that immunoglobulin concentrations in colostrum vary between bitches, with the type of birth significantly influencing the levels. Progesterone withdrawal before birth could play a decisive role in increasing IgG and IgA concentrations in the mammary glands of the bitches.

Impact of Type of Parturition on Colostrum Microbiota Composition and Puppy Survival.

The objective of our study was to determine whether the type of parturition affects the microbiota of the colostrum and the growth and survival of the puppies. Seventy-nine newborn puppies were divided into three groups regarding the type of parturition: vaginal delivery (VD), elective caesarean section (EL-CS), and emergency caesarean section (EM-CS). After the birth of the puppies, swabs of meconium were collected from the puppies and colostrum was obtained from the dam. Many aerobic and anaerobic bacteria were isolated and identified by mass spectrometry (MALDI-TOF MS). The colostrum microbiota of VD and EL-CS puppies contained a significantly higher abundance of bacteria belonging to the genera Staphylococcus, Kocuria and Enterococcus compared with EM-CS colostrum samples. The composition of the meconium microbiota of the puppies present at birth was similar to the colostrum microbiota of their mothers. It was also found that puppies without a meconium microbiota at birth gained weight more slowly compared with puppies with a meconium microbiota at birth. The type of parturition influenced the bacterial composition of the microbiota in the colostrum. Future studies are necessary to further define the significance of the observed differences in microbiota composition between EM-CS compared with EL-CS and VD colostrum microbiota.

Macro- and microelements in serum and seminal plasma as biomarkers for bull sperm cryotolerance.

BACKGROUND: Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality. RESULTS: Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics. CONCLUSIONS: Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance.

Treatment of canine cognitive dysfunction with novel butyrylcholinesterase inhibitor.

Canine cognitive dysfunction (CCD) is common in aged dogs and has many similarities with Alzheimer's disease. Unfortunately, like Alzheimer's disease, CCD cannot be cured. In the present study, we treated dogs with CCD with our newly developed and characterized butyrylcholinesterase inhibitor (BChEi). Seventeen dogs were randomized into two groups (treated with BChEi and untreated) and followed for 6 months at regular check-ups. The dogs' cognitive status was determined by a Canine Dementia Scale (CADES) questionnaire and two cognitive tests. In dogs with moderate cognitive impairment, treatment caused significant improvement in the clinical rating of cognitive abilities and the performance-based tests of cognitive functioning when compared to the untreated group (p < 0.001). Dogs treated with BChEi showed markedly improved cognitive function with enhanced quality of life. No side effects were observed in the treated dogs with moderate cognitive impairment. According to the results of this preliminary study, there is an indication that novel BChEi may be a promising drug for the treatment of CCD in dogs and may be an interesting candidate for the treatment of Alzheimer's disease in humans. However, further clinical studies are needed to confirm this.

Do newborn puppies have their own microbiota at birth? Influence of type of birth on newborn puppy microbiota.

With recent research in humans, a hypothesis known as the sterile womb paradigm has been challenged. The objectives of this study were to determine the presence of placental and fetal microbiomes in dogs, the effect of different types of parturition on the fetal microbiome, and the effect that the fetal microbiome has on early puppy development. A total of 96 newborn puppies from 17 dams were included in the study. Puppies were divided into two groups depending on the type of parturition (vaginal birth (VB) or cesarean section (CS)). Immediately after birth, swabs of the placenta and meconium were taken. Swabs of the oral and vaginal mucosa of the dam were taken in the second half of the pregnancy and just before parturition. All samples were analyzed with a classical bacteriological examination, and bacterial colonies were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The weight gain of each puppy was tracked daily in the first 7 days postpartum. Bacteria from several different genera were isolated from 86.5% of meconium samples and 57% of placenta samples. While the meconium microbiota resembled bacteria from the maternal vagina in VB puppies, the meconium microbiota of puppies born by CS indicated a relative resemblance to maternal oral and vaginal microbiota. A statistically significant difference in the relative growth rate between puppies born by VB and CS was found (p < 0.05), with puppies born by VB gaining weight faster compared to the CS group. This difference was even more noticeable when VB puppies were compared to puppies born by elective CS. Puppies born without a detectable meconium or placental microbiota showed a slower growth rate than those with a meconium microbiota, regardless of the type of parturition (p < 0.05). The findings of this study provide new information about the placental microbiome in healthy pregnant dams and suggest intrauterine colonization of the fetus in dogs. It seems that the type of delivery and bacterial colonization might be an important consideration for the weight gain in puppies in the first few days of life.

Nitrosative Stress in the Frontal Cortex From Dogs With Canine Cognitive Dysfunction.

Canine cognitive dysfunction (CCD) is an age-related disorder similar to human Alzheimer's disease (AD) that occurs in elderly dogs. Nitrosative stress has been implicated as one of the causes leading to neurodegenerative diseases, particularly AD. Its involvement in the development of CCD has not been studied so far. In the present study, immunohistochemical staining detected all three isoforms of nitric oxide synthases (nNOS, eNOS, and iNOS) and 3-nitrotyrosine (3-NT) in brains from CCD-affected dogs and non-demented control dogs in all layers of the canine frontal cortex. In CCD-affected and non-demented brains, nNOS was highly expressed in pyramidal-like neurons in the upper cortical layers. nNOS has also been observed in astrocytes in the CCD frontal cortex. The nNOS immunohistochemical staining was statistically significantly elevated in dogs with CCD in comparison to non-demented dogs. Blood vessel wall cells were positive for eNOS, which was also expressed in astrocytes and neurons. Intense 3-NT immunoreactivity was observed in the upper cortical layers, where amyloid-beta deposits spread in the last stage of CCD. Brain cells in the same area were highly immunoreactive for iNOS. This infers that neuroinflammation and nitrosative stress might exacerbate the neurodegenerative process in CCD-affected brains, ultimately leading to cognitive impairment.

Vitrification Using Soy Lecithin and Sucrose: A New Way to Store the Sperm for the Preservation of Canine Reproductive Function.

A challenge in freezing semen for short and long-term availability is avoiding damage to intact spermatozoa caused by the freezing process. Vitrification protocols provide better results through less manipulation of semen and shorter freezing time compared to slow freezing techniques. Our research was aimed at improving vitrification methods for canine semen. Semen quality was determined in 20 ejaculates after collection. Each ejaculate was divided into eight aliquots, each with a different extender. The control extender contained TRIS, citric acid, fructose, and antibiotics. Soy lecithin and sucrose were added to the control extender at different concentrations to make up the test extenders and final concentration of 50 × 106 spermatozoa/mL. From each group, a 33µL (1.65 × 106 spermatozoa) suspension of spermatozoa was dropped directly into liquid nitrogen and devitrified at least one week later and evaluated as before. Soy lecithin at 1% and 0.25 M sucrose added to the base vitrification media effectively preserved all sperm qualities. Our results demonstrate the effectiveness of our methods. Vitrification media containing sucrose and soy lecithin cause a minimal decline in quality of canine semen after devitrification. Furthermore, extenders used in our research did not contain egg yolk, which was replaced by soy lecithin, thus allowing for ease of shipping to other countries with strict requirements.

Congenital peritoneopericardial diaphragmatic hernia in a family of Persian cats.

CASE SERIES SUMMARY: The congenital midline defects of peritoneopericardial diaphragmatic hernia (PPDH) and omphalocele are believed to be related developmental defects, and have both been described in cats and dogs. This case series describes multiple cases of PPDH and omphalocele in related cats. The majority of cats affected with midline defects (PPDH or omphalocele) were male, consistent with previous reports of male overrepresentation. RELEVANCE AND NOVEL INFORMATION: This is the first report of a family of Persian cats affected by PPDH and/or omphalocele. Clinical findings and pedigree information are suggestive of an autosomal recessive mode of inheritance. However, other modes of inheritance cannot be ruled out owing to limited sample size. The findings in this family and previous reports of Persians affected by PPDH warrant further investigation. Based on this information, recommendations for breeding populations of Persians and long-haired cats should include thoracic radiographs to screen for PPDH until a DNA-based genetic test is available.

Superoxide dismutase: a predicting factor for boar semen characteristics for short-term preservation.

Superoxide dismutase (SOD), total antioxidant capacity (TAC), and thiobarbituric acid reactive substances (TBARS) in seminal plasma were evaluated on the basis of receiver operating characteristics (ROC) analysis as predictors for distinguishing satisfactory from unsatisfactory boar semen samples after storage. SOD on day 0 correlated significantly with progressive motility (r=-0.686; P<0.05) and viability (r=-0.513; P<0.05) after storage; TBARS correlated only with motility (r=-0.480; P<0.05). Semen samples that, after 3 days of storage, fulfilled all criteria for semen characteristics (viability>85%, motility>70%, progressive motility>25%, and normal morphology>50%) had significantly lower SOD levels on the day 0 than those with at least one criterion not fulfilled (P<0.05) following storage. SOD levels of less than 1.05 U/mL predicted with 87.5% accuracy that fresh semen will suit the requirements for satisfactory semen characteristics after storage, while semen with SOD levels higher than 1.05 U/mL will not fulfill with 100% accuracy at least one semen characteristic after storage. These results support the proposal that SOD in fresh boar semen can be used as a predictor of semen quality after storage.