Aerobic Vaginal Microflora in Gestational and Non-Gestational Bitches (Canis lupus familiaris).

The vaginal tract comprises commensal microorganisms, which play an essential role in the health of the reproductive tract. Any dysbiosis in the vaginal microenvironment may lead to severe urinary tract infections or even infertility. This study aimed to evaluate the aerobic bacterial flora isolated from vaginal samples from 100 lactating bitches in the antepartum period (n = 3), postpartum period (n = 80), and with Lactatio sine graviditate (n = 17). Before vaginal swabs, all the bitches went through a gynecology consult, along with milk and blood sampling. Standard microbiological techniques were used for bacterial isolation. Among the 100 vaginal samples analyzed, 82% had a positive microbiological outcome, while 18% were negative. The microbiologic profile listed 17 different genera. The main isolated bacterial families were Micrococcaceae, Staphylococcaceae, Morganellaceae, Bacillaceae, and Rhizobiaceae. At the same time, strains like Agrobacterium radiobacter, Ochrobactrum anthropi, Chromobacterium violaceum, Burkholderia mallei, Bacillus pumllus, or Staphylococcus xylosus were isolated for the first time from the vaginal secretion of lactating bitches. The microbiological data demonstrates that lactating bitches' vaginal discharge is heterogeneous and may be affected by coitus, sampling season, age, and reproductive status.

Interleukin-6 as a Milk Marker of Clinical and Subclinical Intramammary Infections (IMI) in Cows Caused by Streptococcus spp.

The aim of the study was to evaluate the concentrations of Interleukin-6 (IL-6) in milk and serum of healthy cows (HE) and cows with mastitis caused by Streptococcus spp. The blood and milk samples were obtained from Holstein-Friesian cows (Lublin region, Poland). A total of 43 milk and serum samples from 28 cows with mastitis and 15 healthy cows were selected for study. IL-6 levels in milk from HE cows ranged from 6.09-80.24 pg/mL (median 26.6 pg/mL) and were significantly lower than in milk from both cows with clinical and subclinical mastitis (487.09 pg/mL vs. 26.6 pg/mL in CM, p < 0.001; and 165.31 pg/mL vs. 26.6 pg/mL in SCM, p < 0.001). The IL-6 concentration in the serum of HE was not significantly different from the serum IL-6 of the entire group of mastitis cows, regardless of whether the inflammation proceeded in a clinical or subclinical form (44.37 pg/mL vs. 78.09 pg/mL; 128.29 pg/mL vs. 78.09 pg/mL, respectively). The present study indicates that cows with mastitis caused by Streptococcus spp. develop a local immune response in the mammary gland in response to the pathogen. Monitoring of IL-6 levels in milk can allow early detection of mastitis, which is especially important in cases of subclinical inflammation.

Evaluation of the presence of gingivitis as confounding factor in assessing inflammatory status in serum and saliva of dogs with diabetes mellitus.

The aim of this study was to evaluate the changes in the serum and salivary inflammatory markers induced by Diabetes mellitus (DM) in dogs and to assess the possible confounding effect of gingivitis. A panel of 13 cytokines was measured in the serum and saliva of dogs diagnosed with DM and compared with healthy dogs without gingivitis (control group 1; CG1) and dogs with gingivitis but otherwise healthy (control group 2; CG2). The results of the present study showed statistically significantly higher levels of IL-8, KC-like and MCP1 in the serum of dogs with DM compared to CG1 dogs. In the case of saliva, the DM group presented statistically higher GM-CSF, IL6, IL15, and MCP1 levels compared to CG1, and lower KC-like chemokine compared to CG2. Finally, gingivitis produced changes in saliva, with salivary levels of GM-CSF, IL-6, IL-7, IL-15, IP-10, KC-like, IL-10, IL-18, MCP1, TNFα being statistically significantly higher in the saliva of CG2 dogs compared to CG1. The results of the present study indicate that dogs with DM have altered cytokine levels in serum and saliva compared to healthy dogs. In addition, this study highlights the importance of taking oral health into account when determining cytokines in dogs, as gingivitis can significantly alter their concentrations. .

Relationships between haptoglobin and inter-alpha trypsin inhibitor heavy chain 4 levels in local and peripheral blood and systemic inflammatory response syndrome in bitches with pyometra.

We aimed to assess the usefulness of monitoring inter-alpha trypsin inhibitor heavy chain 4 (ITIH4) and haptoglobin (Hp) in peripheral and local blood in canine pyometra, and evaluation the relationships among acute phase proteins (APPs), systemic inflammatory response syndrome (SIRS) and the presence of bacteria. The material was collected from bitches with pyometra and from healthy ones. Blood was taken from the cephalic and uterine veins. APPs levels were quantified by ELISA. In the peripheral circulation, the Hp was higher in animals with open-cervix pyometra (OCP) than in the closed-cervix pyometra (CCP) and the control group. The Hp concentration was not correlated with age, with the presence of SIRS or with the type of bacteria (Gram-negative, Gram-positive or mixed flora). The ITIH4 concentrations in the peripheral blood did not differ significantly in the cases of pyometra. The Hp concentration in the local circulation increased in the OCP but not in the CCP groups, although the histopathological changes in the endometrium were similar. Peripheral Hp concentrations may be a useful tool in differentiating between the types of pyometra.

Comparative analysis of total protein, casein, lactose, and fat content in milk of cows suffering from subclinical and clinical mastitis caused by Streptococcus spp.

Introduction: The aim of the study was to analyse the total protein (TP), casein (CAS), lactose (LAC), and fat content of milk from cows with subclinical (SCM) and clinical mastitis (CM) caused by Streptococcus spp. Material and Methods: A total of 60 milk samples from diseased cows and 30 milk samples from healthy cows were included in the study. Milk samples were taken from Holstein-Friesian cows from four dairy farms in Lublin Province. The bacteriological examination of the milk was performed and the somatic cells count in 1 mL of milk was determined using a SomaCount FC automatic cell counter. Determination of TP, CAS, LAC, FAT and FA levels in milk was carried out using a DairySpec FT automated Fourier transform infrared spectrometer. Results: Total protein in milk from HE was significantly higher than in milk from cows with mastitis (4.04% vs 3.57% in milk from SCM cows and 3.7% in milk from CM cows, P = 0.001). The CAS level was 2.73% in milk from CM cows and 2.92% in milk from SCM cows vs 3.30% in milk from HE cows, P = 0.001. The changes in CAS and TP in milk resulted in a significant difference in the CAS/TP ratio (81.7% in milk from HE cows vs 73.8% in milk from CM cows). A decrease in levels was also recorded for LAC (4.8% in milk from HE cows vs 4.51% in milk from SCM cows and 4.01% in milk from CM cows, P = 0.001). The fat level was significantly higher in milk from healthy cows than in milk from cows with mastitis (4.0% vs 2.3% in milk from SCM cows and 1.64% in milk from CM cows, P = 0.001). Conclusion: It should be emphasised that the decrease in the levels of TP, LAC and FAT was significant not only in milk from CM cows but also in milk from SCM cows. This is very unfavourable, because the reduction in the main milk components results in poor quality dairy products and impairs line processes.

Impact of carbon quantum dots on self-assembly and dielectric relaxation modes of a room temperature tri-component fluorinated antiferroelectric liquid crystal mixture.

We investigated the impact of carbon quantum dots (CQDs) of sizes ∼2-5 nm in a room temperature tri-component fluorinated antiferroelectric liquid crystal (AFLC) mixture. The synthesised CQDs have been characterised by high-resolution transmission electron microscopy and X-ray diffraction spectroscopy. An increase in the transition temperature and enthalpy to the isotropic liquid phase indicates stabilization of the AFLC mixture in the presence of CQDs. The dielectric studies have been carried out in the frequency range of 1 Hz-40 MHz under the planar anchoring conditions of the molecules. An appreciable increase in the permittivity, dielectric strength, and conductivity has been observed owing to the existence of sp2/sp3 hybridization in the CQDs which form a strong coupling and develop a dipolar ordering in the systems. Various relaxation frequencies were increased with the incarceration of CQDs in the AFLC mixture. Overall, the different studies suggest that the doped CQDs are very well settled in between the host molecules without disturbing the molecular ordering of the hosts. Such results are encouraging and reveal the potential applicability of the CQD doped systems to produce highly efficient tuneable optical devices and other multifaceted applications.

Gold Nanorod-Induced Effects in a Mesogenic Compound 4-(trans-4-n-Hexylcyclohexyl) isothiocyanatobenzene.

Gold nanorods (GNRs) have a capsule-like structure with different optical properties than spherical gold nanoparticles due to surface plasmon resonance. Liquid crystals (LCs) are mesogenic compounds having crystal-like orientation and liquid-like fluidity. They are important materials from a technological point of view. Both GNRs and LC compounds are anisotropic in shape and properties. Different nano entities show interesting results when dispersed in different liquid crystalline materials which are instrumental from the application point of view. In the present work, GNRs have been dispersed in nematic liquid crystalline materials, namely 4-(trans-4-n-hexylcyclohexyl) isothiocyanatobenzene (6CHBT). Calorimetric, texture, spectroscopic, and dielectric studies were carried out for a pure 6CHBT and its composites with GNRs. Different calorimetric and dielectric parameters such as transition temperature, enthalpy, heat flow, permittivity, dielectric strength, dielectric anisotropy, and relaxation frequency have been determined, and the effect of GNRs has been explored. This article gives an insight into the influence of GNRs on the morphology and anisotropic physical properties of the nematic liquid crystalline material.

Mammary gland inflammation in rabbits does (Oryctolagus cuniculus): A systematic review.

Mastitis is a common condition in rabbit (Oryctolagus cuniculus) does, especially in rabbitries where mammae inflammations are the leading pathological expression of Staphylococcus aureus infections in reproductive females. It affects both the litter and the lactating female, including subsequent lactations. Non-septic (i.e. galactostasis and cystic mastitis) and septic (i.e. Mastitis acuta; including Mastitis gangrenosa; and Mastitis chronica) mammary inflammations are acknowledged; however, the literature is scarce regarding non-septic inflammations or subclinical mastitis. Thus, the present systematic review highlights and summarizes the existing data concerning mastitis in rabbit does, including diagnosis and treatment plans.

A systematic review of mammary gland inflammations in queens (Felis catus).

Mastitis is a rare condition that can be diagnosed in female cats of all breeds and ages. It is usually caused by bacterial agents and is frequently encountered secondarily in feline mammary fibroepithelial hyperplasia (FEH). If left untreated, it can peril both the queen and her kittens. Therefore, a reliable and quick diagnosis is the primary key to successful treatment. Conventional diagnosis consists of a clinical and ultrasound evaluation of the queen's mammary glands. However, there is a lack of literature specifically focused on mastitis in queens. Consequently, the present review brings forth the existing knowledge regarding mammary gland inflammations in cats, with the goal of improving current clinical evaluation and treatment plans.

Divergences between serum C-reactive protein and ferritin concentrations in canine pyometra.

The main aim of this report was to investigate and compare the response of serum C-reactive protein (CRP) and ferritin, two positive acute phase proteins (APPs) which usually show an increase in inflammatory processes, in dogs with pyometra. For this purpose, two different studies were made. In the first one , both proteins were measured together in an APPs profile in 25 dogs with pyometra, 25 dogs with pancreatitis (as an example of a positive inflammatory control group), and in 25 healthy dogs. In the second study, to advance the knowledge of the changes and evolution of serum ferritin and CRP in dogs with pyometra after treatment, the concentrations of both APPs were analyzed in 30 dogs with pyometra at diagnosis and after ovariohysterectomy and in 10 clinically healthy female dogs before and after elective spaying. In both studies, bitches with pyometra showed significant increases in serum CRP, indicating an inflammatory condition, but not in serum ferritin despite being a moderate positive APP. This divergence between the dynamics of these APPs could be a useful tool for the suspicion of cases of canine pyometra.

Enhancement of dielectric and electro-optical characteristics of liquid crystalline material 4'-octyl-4-cyano-biphenyl with dispersed functionalized and nonfunctionalized multiwalled carbon nanotubes.

For recent applications, liquid crystal-carbon nanotube based nanocomposite systems have been proven to be highly attractive. In this paper, we give a thorough analysis of a nanocomposite system made of both functionalized and nonfunctionalized multiwalled carbon nanotubes that are disseminated in a 4'-octyl-4-cyano-biphenyl liquid crystal medium. Thermodynamic study reveals a decrease in the nanocomposites' transition temperatures. In contrast to nonfunctionalized multiwalled carbon nanotube dispersed systems, the enthalpy of functionalized multiwalled carbon nanotube dispersed systems has increased. In comparison to the pure sample, the dispersed nanocomposites have a smaller optical band gap. A rise in the longitudinal component of permittivity and, consequently, the dielectric anisotropy of the dispersed nanocomposites has been observed by dielectric studies. When compared to the pure sample, the conductivity of both dispersed nanocomposite materials has increased by two orders of magnitude. For the system with dispersed functionalized multiwalled carbon nanotubes, the threshold voltage, splay elastic constant, and rotational viscosity all decreased. For the dispersed nanocomposite of nonfunctionalized multiwalled carbon nanotubes, the value of the threshold voltage is somewhat decreased but the rotational viscosity and splay elastic constant both are enhanced. These findings show the applicability of the liquid crystal nanocomposites for display and electro-optical systems with appropriate tuning of the parameters.

Assessment of Selected Immunological Parameters in Dairy Cows with Naturally Occurring Mycotoxicosis before and after the Application of a Mycotoxin Deactivator.

Introduction: Mycotoxins in dairy cows can cause many non-specific symptoms often resulting from immune system overreaction. The study assessed the concentration of selected cytokines and acute phase proteins (APP) in cows with natural mycotoxicosis before and after using a mycotoxin neutraliser. The cytokines were tumour necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and interleukin 10 (IL-10), and the APP were serum amyloid A (SAA) and haptoglobin (Hp). Material and Methods: The research was carried out on an experimental group (Exp) of 10 herdmate Holstein-Friesian cows with mycotoxicosis. The control group (Con) was 10 healthy cows of the same breed from a different herd. Cows in the Exp group were administered the mycotoxin deactivator Mycofix for three months. Blood was drawn from Exp cows once before administering Mycofix and a second time after three months of its use. Blood was also drawn from Con cows at the same times. Serum levels of TNF-α, IL-6, IL-10, SAA and Hp were assessed using ELISA. Results: The concentrations of all cytokines and Hp in Exp cows were higher before treatment (P < 0.001) than those in Con cows. After three months of administering Mycofix, the concentrations of TNF-α and IL-6 were significantly lower than their pre-treatment levels (P < 0.001). The concentrations of IL-6, IL-10, and Hp were still significantly higher than those in the Con group (P < 0.001). In cows with mycotoxicosis, simultaneous stimulation of antagonistic processes was noted: a pro-inflammatory process in the upregulation of TNF-α and IL-6, and an anti-inflammatory one in the upregulation of IL-10. Conclusion: Despite the absorbent's use and the resolution of clinical symptoms in Exp cows, high levels of IL-10 and Hp and IL-6 were maintained. Assessment of the level of cytokines and APP appears to be a useful and precise tool for the evaluation and application of the appropriate dose of the mycotoxin absorbent or the evaluation of its effectiveness.

Phagocytic Activity, Oxygen Metabolism and Serum Amyloid a Concentration in Peripheral Blood of Mink with Subclinical Aleutian Virus Infection.

Aleutian disease (AD) is a chronic disease of mink caused by the Aleutian Mink Disease Virus (AMDV) that results in dysfunction of the immune system. The prevalence of asymptomatic AMDV infections suggests a necessity to explore their effects on the cellular mechanisms of non-specific immunity in farmed mink. The study evaluated the phagocytic activity and oxygen metabolism of peripheral blood granulocytes and monocytes in mink with chronic subclinical AMDV infection. Moreover, the intensity of inflammatory processes was assessed based on the serum amyloid A (SAA) concentration. The analyses involved 24 brown mink females aged 12−24 months. The experimental group (group I) consisted of mink with chronic subclinical AMDV infections, and the control group (group II) included healthy animals. The statistical analysis was performed using the Mann-Whitney U rank test. Phagocytic activity of granulocytes and monocytes was carried out using flow cytometry, and SAA concentration was determined with enzyme-linked immunosorbent assay (ELISA). Compared with the control group, there was a significant decrease in the phagocytic activity and oxygen metabolism of granulocytes and monocytes in the AMDV-infected mink (p < 0.05). Additionally, it was found that the mean SAA value was significantly higher in the group infected with AMDV than in the control group (p < 0.05). The obtained data indicate that monitoring the serum SAA levels in mink with asymptomatic inflammation may help assess the health of mink and detect asymptomatic inflammation caused by AMDV infection.

Activity of the glycosidases ß-galactosidase, α-l-fucosidase, ß-N-acetyl-hexosaminidase, and sialidase in uterine tissues from female dogs in diestrus with and without pyometra.

This study aimed to compare the activity of selected glycosidases (ß-galactosidase, α-l-fucosidase, ß-N-acetyl-hexosaminidase, and sialidase) in homogenates of uterine tissues obtained from female dogs with and without pyometra. In addition, it examined the availability of substrates for these glycosidases in the homogenates. The study was carried out on female dogs undergoing ovariohysterectomy for pyometra (n = 10) and clinically healthy dogs (n = 10) undergoing elective spaying. The activity of ß-galactosidase, α-l-fucosidase, and ß-N-acetyl-hexosaminidase was analyzed using a spectrofluorometer and that of sialidase using a colorimetric method. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis with Alcian Blue (AB) and Periodic Acid-Schiff (PAS) staining was performed to determine the presence of substrates for these glycosidases in the homogenates of uterine tissues. The results revealed that the activity of all the examined glycosidases was significantly higher (P < 0.05) in the uterine tissues isolated from dogs with pyometra in comparison to healthy dogs. The electrophoretic patterns of the selected samples showed several proteins, which contained different sugar moieties stained by AB and PAS and the profiles differed significantly between the pyometra group and the healthy group. Densitometric analysis of AB staining showed patterns between 233 and 148, 86 and 55, and 43 and 20 kDa, which differed markedly in sugar content between the examined groups of animals. Similarly, PAS staining analysis revealed patterns of different molecular weights, between 233 and 117 and between 55 and 32 kDa, which also differed in sugar content. These findings suggest that canine pyometra is accompanied by the increase in the activity of selected glycosidases in the uterus. This could potentially modify the glycan structures of uterine glycoproteins and in result their biological functions. Further studies are needed to elucidate the potential role of the increased activity of glycosidases in the pathogenesis of this disease.

Content of tryptophan and kynurenines in serum and milk of dairy cows with mastitis caused by Streptococcus spp.

The aim of the study was to investigate serum and milk concentrations of tryptophan (TRP), kynurenine (KYN) and kynurenic acid (KYNA), and activity of indoleamine 2,3-dioxygenase (IDO) in cows suffering from mastitis caused by Streptococcus spp. The blood and milk samples were collected from Holstein-Friesian cows farmed in the Lublin region of Poland. It was found that TRP was lower in cows with mastitis both in serum and milk. KYN was lower in serum but not in milk. KYNA was not significantly altered in diseased cows both in serum and milk. The activity of IDO calculated as KYN to TRP ratio was unchanged in serum but was markedly elevated in milk of cows with mastitis. Our findings may have important implications for diagnosis of mastitis in cows because an increase of activity of IDO and reduction of TRP in milk might be a valuable early marker predicting the occurrence of the disease.

Aerobic Isolates from Gestational and Non-Gestational Lactating Bitches (Canis lupus familiaris).

Mastitis is a complex and well-defined mammary gland pathology, and an emergency in bitches. In dogs, its prevalence is about 1% of all reported diseases and about 5.3% of all reproductive pathologies. Lactating bitches are naturally prone to developing mastitis since puppies can easily overstimulate the epidermal layer of nipples during feeding, facilitating bacterial colonization of the glands. This study aimed to describe the aerobic bacterial flora isolated from milk samples derived from a cohort of patients (n = 87) diagnosed with clinical mastitis (n = 29), subclinical mastitis (n = 17) and healthy mammary glands (n = 46). All of the patients underwent a gynecology consultation to diagnose mammary gland afflictions; physical examination results were coupled with traditional hematological findings. The milk samples were plated on specific microbiological media for bacterial isolation. Among the 162 milk samples analyzed, 93.2% (151/162) had a positive microbiological result, while 6.8% (11/162) were sterile. The bacteriological profile of the milk samples showed 47 different species. The most common bacterial families detected in healthy bitches and bitches with subclinical and clinical mastitis were the Staphylococcaceae, Enterobacteriaceae and Enterococcaceae families. The results indicated that half of the isolated bacteria are novel findings in dogs and that some of them are normal components of human milk.

MicroRNA and vascular endothelial growth factor (VEGF) as new useful markers in the diagnosis of benign prostatic hyperplasia in dogs.

Numerous specific biomarkers with a prognostic and diagnostic value comparable to histopathological findings are now used for non-invasive diagnosis of prostate diseases in humans. Meanwhile, as far as dogs are concerned, the diagnosis of prostate disorders is based solely on clinical examination and ultrasound (USG). Therefore, the aim of the study was to assess the usefulness of two biomarkers, i.e. miRNA-129 and VEGF for the diagnosis of BPH in dogs. The study involved 40 dogs divided into three groups. Group I (n = 9) comprised healthy dogs up to the age of 5 years, Group II (n = 17) comprised dogs between the ages of 5-10 suffering from BPH as confirmed by the examination and Group III (n = 14) comprising dogs over 10 years of age, which also had BPH confirmed. The results demonstrated that dogs in group II and III exhibited a significant decrease in miRNA expression (P < 0.0001) and a significant increase in serum VEGF levels (P = 0.025) when compared to the dogs in group I. There was also a positive correlation between the prostate size and VEGF level. The findings led to the conclusion that the determination of miRNA-129 and VEGF can significantly contribute to the diagnosis of prostate disorders in dogs.

Measurement of ITIH4 and Hp levels in bitches with pyometra using newly developed ELISA methods.

Measurement of acute phase proteins (APPs) as biomarkers in canine medicine is in increasing demand. In the present study, the development and validation of two ELISA methods for the quantification of canine inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4) and haptoglobin (Hp) are shown. The adequate imprecision and accuracy and wide analytical range make the developed methods appropriate to quantify ITIH4 and Hp in serum samples. The inter- and intra-assay CVs were lower than 10 %, and the assays maintained linearity under dilution and showed analytical equivalence with the method of radial immunodiffusion. The measurement of CRP, Hp and ITIH4 in sera from bitches affected by pyometra allowed us to determine that ITIH4 behaves as a moderate APP in dogs. The group of bitches affected by pyometra showed very high levels of CRP (147 ± 91 mg/L), corresponding to a strong inflammatory process, which resulted in a moderate increase in the concentrations of Hp (7 times) and ITIH4 (3 times) compared to the control group.

Changes in salivary proteins can reflect beneficial physiological effects of ejaculation in the dog.

The objective of this study was to study the changes in salivary proteins that occur in the dog after the ejaculation process. Saliva samples from eight dogs before and after induced ejaculation were analyzed by proteomic using Tandem Mass Tag (TMT) labeling and LC-MS/MS analysis. A total of 33 salivary proteins showed significant changes after the ejaculation process. The up-regulated proteins that showed changes of higher magnitude were mucin-7 (MUC-7), peroxiredoxin-4 (PRDX4) and galectin-3 (LEGALS3) whereas proteins such as alpha-1-acid glycoprotein (A1G1) and alpha-1B-glycoprotein (A1BG) were the most down-regulated. MUC-7 and PRDX4 expression in saliva after ejaculation could be associated with the protective "environment" created by the organism to exert pr 3o-fertility activities and antioxidants benefits in spermatozoa. Also LEGALS3 increment could be associated with an improvement of wellbeing and could contribute to a positive global effect in the body. Down-regulations of A1G1 and A1GB proteins found in saliva after ejaculation could be associated with a reduction in systemic inflammation. Overall it can be concluded that, changes in proteins in saliva that are produced after ejaculation can reflect a state of increase immune defenses, improvement of antioxidant status and low inflammation.

Lactation-related mammary gland pathologies-A neglected emergency in the bitch.

Mastitis is a common reproductive disorder in bitches, reaching a prevalence of 0.71%. Mastitis has a wide range of forms, from asymptomatic to severe gangrenous mastitis that can lead to septic shock and death of the bitch and nurslings. However, most of the time it is overlooked, undiagnosed or mistreated. The present systematic review was performed to revise and summarize the existing knowledge related to this disorder, including diagnosis, treatment and prevention.