Bacterial Contamination of the Surgical Site at the Time of Elective Caesarean Section in Belgian Blue Cows-Part 2: Identified by 16Sr DNA Amplicon Sequencing.

This study aimed to describe the bacterial composition of the surgical site during elective caesarean sections (CSs) using the 16Sr DNA amplicon sequencing performed in parallel to bacterial culture. The study involved 13 Belgian blue cows of a previous dataset of 76 animals. Bacteriology was performed on swabs sampled from visceral and parietal peritoneum during the CS. Amplicon sequencing was performed in six samples chosen randomly among the swabs positive for bacteriology and seven among the culture-negative swabs. A total of 2542 bacterial operational taxonomic units belonging to 567 genera were identified. The most often identified genus and species were Mycoplasma (44%) and Mycoplasma wenyonii (36%), respectively. Results showed no difference in microbiota composition between the culture-positive and -negative samples. However, a difference was observed between the bacteriology and amplicon sequencing results. Indeed, seven out of nine cultured strains were not identified by amplicon sequencing in the samples in which they were cultured. In contrast to bacteriology, amplicon sequencing unveiled the presence of bacterial DNA in all elective CSs. The most identified DNA is most likely derived from the haematogenous spread of bacteria to the surgical site. Furthermore, the cultured bacteria were not the dominant species in the sample from which they were cultured.

Bacterial Contamination of the Surgical Site at the Time of Elective Caesarean Section in Belgian Blue Cows-Part 1: Identified by Bacterial Culture.

To improve the efficacy of preoperative antibiotics used in elective caesarean section (CS), we aimed to identify the bacteria contaminating the surgical site during this surgery. A study was conducted on 76 Belgian Blue cows. Bacteriology was performed on cotton swab sampled from the visceral and parietal peritoneum of each cow during the CS. Most of samples showed a negative culture (55/76; 72.37%), 19/76 (25%) were positive (p < 0.0001) and two samples were contaminated. In total, 32 isolates belonging to 18 species were identified. Most of them are aerobic (17/18; 94.44%) and half of them were gram-negative (G-). The most encountered bacteria were Acinetobacter sp. (6/32; 18.75%), Pseudomonas sp. (4/32; 12.5%), Aerococcus viridans (4/32; 12.5%), Psychrobacter sp. (3/32; 9.37%), and Escherichia coli (2/32; 6.25%). Among the identified isolates, 31/32 (96.87%) were aerobic and 1/32 (3.12%) was anaerobic (p = 0.0001). Furthermore, 20/32 (62.50%) strains were G− while 12/32 (37.5%) were gram-positive (G+) (p = 0.012). In fact, most of cultured strains were aerobic G− (20/32), 11/32 were aerobic G+ and 1/32 is anaerobic G+ (p < 0.0001). In conclusion, most of samples showed a negative bacteriology; however, aerobic G− strains were the most identified in positive swabs. Therefore, preoperative antibiotics should be aimed against these bacteria.

Blood Inflammatory, Hydro-Electrolytes and Acid-Base Changes in Belgian Blue Cows Developing Parietal Fibrinous Peritonitis or Generalised Peritonitis after Caesarean Section.

This study aimed to describe the inflammation, hydro-electrolyte and acid-base imbalances caused by generalised peritonitis (GP) and parietal fibrinous peritonitis (PFP) after caesarean section. After clinical examination, blood was sampled from 11 cows with PFP, 30 with GP and 14 healthy cows. Serum and plasma refractometry and glutaraldehyde tests were used to evaluate the inflammation level, while hydro-electrolytes and acid-base parameters were assessed using an EPOC® device. In addition to clinical signs of dehydration (>10%), blood analysis showed a high fibrinogen concentration (PFP: 8.64 ± 8.82 g/L; GP: 7.83 ± 2.45 g/L) and fast glutaraldehyde coagulation (<3 min) indicative of severe inflammation in both diseases compared to the control group (p < 0.05). Moreover, a severe decrease in electrolytes concentration (Na+: 126.93 ± 5.79 mmol/L; K+: 3.7 ± 1.3 mmol/L; Ca++: 0.89 ± 0.12 mmol/L; Cl−: 82.38 ± 6.45 mmol/L) and a significant increase in bicarbonate (30.87 ± 8.16 mmol/L), base excess (5.71 ± 7.42 mmol/l), L-lactate (8.1 ± 4.85 mmol/L) and creatinine (3.53 ± 2.30 mg/dL) were observed in cows with GP compared to the control group (p < 0.05). In contrast, few major perturbations were noticed in PFP, where only K+ (3.64 ± 0.25 mmol/L) and Ca++ (1.06 ± 0.09 mmol/L) were significantly modified (p < 0.05). In conclusion, a high dehydration and severe inflammation are induced by PFP and GP. Nevertheless, GP causes more electrolytes and acid-base disturbances than PFP.

Antimicrobial Susceptibility Profile of Several Bacteria Species Identified in the Peritoneal Exudate of Cows Affected by Parietal Fibrinous Peritonitis after Caesarean Section.

The aim of this study was to identify the species and antimicrobial susceptibility of bacteria involved in parietal fibrinous peritonitis (PFP). We studied 156 peritoneal fluid samples from cows presenting PFP after caesarean section. Bacteria were cultured in selective media and their antimicrobial susceptibility was tested by disk diffusion assay. Bacteria were isolated in the majority (129/156; 83%) of samples. The majority (82/129; 63%) of positive samples contained one dominant species, while two or more species were cultured in 47/129 (36%) samples. Trueperella pyogenes (T. Pyogenes) (107 strains) was the most identified species, followed by Escherichia coli (E. coli) (38 strains), Proteus mirabilis (P. mirabilis) (6 strains), and Clostridium perfringens (C. perfringens) (6 strains). Several other species were sporadically identified. Antimicrobial susceptibility was tested in 59/185 strains, predominantly E. coli (38 strains) and P. mirabilis (6 strains). Antibiotic resistance, including resistance to molecules of critical importance, was commonly observed; strains were classified as weakly drug resistant (22/59; 37%), multidrug resistant (24/59; 41%), extensively drug resistant (12/59; 20%), or pan-drug resistant (1/59; 2%). In conclusion, extensive antibiotic resistance in the isolated germs might contribute to treatment failure. Ideally, antimicrobial therapy of PFP should be based upon bacterial culture and susceptibility testing.

Description of Plasma Penicillin G Concentrations after Intramuscular Injection in Double-Muscled Cows to Optimize the Timing of Antibiotherapy for Caesarean Section.

In order to improve the efficacy of penicillin injection during caesarean section, we aimed to identify the optimal timing of its preoperative administration. A study was conducted in 12 adult, non-pregnant Belgian Blue cows. To evaluate the plasma penicillin concentrations, blood samples were taken from the jugular vein at -5, 15, 30, 45, 60, 120, 240, 480 min relative to the intramuscular (IM) injection of 21,000 IU/kg of body weight of penicillin G. Results showed that plasma concentrations at 15 min after IM injection (668.3 ± 73.7 ng/mL) largely exceeded the minimal inhibitory concentration (MIC) of penicillin-sensitive bacteria (MIC < 125 ng/mL). With increasing time, plasma concentrations continued to rise, attaining an increasing proportion of moderately sensitive bacteria (250 ng/mL > MIC < 2000 ng/mL). The maximal concentration was reached between 1 and 4 h (average: 1.495.1 ± 181.7 ng/mL) after IM injection in the majority of cows, and decreased non-significantly to 1002.1 ± 93.2 ng/mL at 8 h. In conclusion, plasma penicillin concentrations at 15 min after an IM injection inhibit penicillin-sensitive bacteria. However, in order to obtain the maximal protective effect of the antibiotherapy, surgery should be started at 1 to 2 h after IM penicillin injection.

Parietal fibrinous peritonitis in cattle: A literature review.

BACKGROUND: Parietal fibrinous peritonitis (PFP) is a complication of laparotomy in cattle, consisting of fluid and fibrin accumulation within a fibrous capsule between the parietal peritoneum and the abdominal muscles. Since scientific information on PFP is scarce, we aim to collect available information to help practitioners in its diagnosis and treatment, and to formulate research perspectives. METHODS: PubMed and GoogleScholar databases were scanned using "cattle" or "bovine", and one of the following keywords: "seroma", "parietal fibrinous peritonitis", "retroperitoneal abscess", or "wound infection". RESULTS: Although scientific information is often anecdotal, two recent larger studies shed more light on PFP symptoms, diagnosis and treatment. Symptoms vary according to the cavity's localisation and size, and include anorexia, weight loss and an inflammatory status. Rectal palpation is strongly indicative, but the definitive diagnosis is made by ultrasound. Trueperella pyogenes and Escherichia coli are frequently isolated germs, although it remains unclear whether they are primary or secondary agents. Good survival rates were reported after surgical drainage. CONCLUSION: Although the diagnosis and treatment seem clear, the exact pathogenesis of PFP should be the focus of ongoing research. This can be achieved by epidemiological data analysis focusing on risk factors like surgery technique, housing and ration.

Treatment protocols and management of retained fetal membranes in cattle by rural practitioners in Belgium.

Retained fetal membranes (RFM) is a frequent postpartum disorder in cattle causing considerable economic losses, and a common indication for antibiotic (AB) administration. There is controversy with regard to the treatment of RFM, and scientific recommendations are often in conflict with current legislation on AB use and practical routines in the field. The aim of this study was to assess the therapeutic approaches of RFM by Belgian rural veterinarians. A digital survey was sent to 468 Belgian veterinarians; 149 complete questionnaires (Wallonia: 78; Flanders: 71) were obtained. Survey questions captured socio-demographic characteristics, case definition, therapeutic approaches and treatment options for RFM. Questions on treatment were duplicated for dairy and beef cattle. When confronted with dairy cows suffering from RFM without fever, 35.6 % of vets do not treat with AB, while the majority administers AB, either intrauterine (47.6 %), systemically (10.7 %) or both (6.1 %). Dairy cows with RFM and fever receive a systemic (33.5 %), intrauterine (2.7 %) or combined (61.8 %) AB treatment. For a beef cow with RFM without fever, 21.5 % of vets prefer no AB treatment, while others prefer an intrauterine (24.2 %), systemic (24.8 %), combined (29.5 %) AB treatment. Beef cows with RFM and fever receive AB from the large majority of vets, by systemic (34.9 %) or combined (56.3 %) administration. In case of a parenteral treatment, benzylpenicillin, amoxicillin and ampicillin are by far the most frequently used molecules; only a minority of vets use cephalosporins. Manual placental removal is attempted by 93.9 % of the respondents. These results demonstrate that there is room for improvement when considering the treatment protocols of RFM. Many veterinarians use AB in RFM cows not presenting symptoms of general illness, or administer intrauterine AB in cows with or without clinical signs of illness. Concerning the molecules of choice, practical routines are largely in line with national AB recommendations. Beef cows suffering from RFM are treated more rigorously than dairy cows, regardless of their rectal temperature.

Infectious Agents Identified by Real-Time PCR, Serology and Bacteriology in Blood and Peritoneal Exudate Samples of Cows Affected by Parietal Fibrinous Peritonitis after Caesarean Section.

The aim of this study was to identify the pathogens potentially involved in parietal fibrinous peritonitis (PFP). PFP is a complication of laparotomy in cattle, characterized by an accumulation of exudate inside a fibrinous capsule. We have studied 72 cases of PFP in Belgian blue cows, confirmed by a standard diagnostic protocol. Blood was collected to evaluate the presence of antibodies for Mycoplasma bovis(M. bovis), Coxiella burnetii(C. burnetii) and Bovine Herpesvirus 4(BoHV4) by enzyme-linked immunosorbent assays. Peritoneal exudate was obtained from the PFP cavity to perform bacteriological culture, and to identify the DNA of M. bovis, C. burnetii and BoHV4 using real time polymerase chain reaction (qPCR). Bacteriological culture was positive in most peritoneal samples (59/72); Trueperella pyogenes (T. pyogenes) (51/72) and Escherichia coli (E. coli) (20/72) were the most frequently identified. For BoHV4, the majority of cows showed positive serology and qPCR (56/72 and 49/72, respectively). Contrariwise, M. bovis (17/72 and 6/72, respectively) and C. burnetii (15/72 and 6/72, respectively) were less frequently detected (p < 0.0001). Our study proves that PFP can no longer be qualified as a sterile inflammation. Moreover, we herein describe the first identification of BoHV4 and C. burnetii in cows affected by PFP.

Comparison between generalised peritonitis and parietal fibrinous peritonitis in cows after caesarean section.

BACKGROUND: Parietal fibrinous peritonitis (PFP) and generalised peritonitis (GP) are two postoperative complications in cows, characterised by fluid and fibrin accumulation throughout the peritoneum (GP) or in an encapsulated cavity (PFP). Unlike GP, PFP is scarcely documented. METHODS: Twenty-one GP cases and 12 PFP cases were confirmed by ultrasound in cows referred to the Veterinary Clinic (Liège University) for complications after caesarean section. All cows underwent a standardised examination protocol. Blood samples were analysed for metabolic and inflammatory markers. Bacteriology was performed on peritoneal fluid samples. Treatment consisted of surgical drainage of the abdominal cavity (GP) or the encapsulated cavity (PFP). Variables concerning anamnesis, clinical findings and treatment outcomes were compared. RESULTS: Perioperative complications had occurred in 9/21 GP cows but 0/12 PFP cows (P<0.05). Biochemical analysis indicated pronounced inflammation and did not differ between groups. Peritoneal fluid samples of both groups were contaminated and contained similar bacteria (Trueperella pyogenes and Escherichia coli). While 11/12 PFP cows were discharged, all patients with GP died or were euthanased (P<0.05). CONCLUSIONS: We hypothesise that PFP and GP are two different manifestations of perioperative peritoneal contamination. The severity and spread of the contamination determine the clinical presentation and the prognosis.

Prophylactic antibiotic usage by Belgian veterinarians during elective caesarean section in Belgian blue cattle.

The use of prophylactic antibiotics in veterinary surgery reduces the occurrence of postoperative complications. In order to limit the development of antimicrobial resistance in bacteria and to maximize therapy efficacy, antibiotics should be used prudently and efficiently. A survey was sent to Wallonian rural veterinarians in order to assess the use of antibiotics before, during and after bovine elective caesarean section, the most common surgical procedure in Belgian rural practice. Results were analysed in the light of the current guidelines formulated by the Belgian Centre of Expertise on Antimicrobial Consumption and Resistance in Animals. Among 380 contacted veterinarians, 113 answered the survey. All veterinarians use antibiotics during caesarean section. Veterinarians predominantly use penicillin as the first drug of choice, whereas a minority use drugs considered as second or third choice, such as amoxicillin, oxytetracyclin or lincomycin-spectinomycin. Also, 31/113 veterinarians simultaneously use molecules from different antibiotic classes. The majority (100/113) of veterinarians inject antibiotics during or after surgery, while a minority (13/113) administer antibiotics preoperatively. Most veterinarians (67/113) limit the duration of their antibiotic treatment to 1 day. Considering the administration route, most veterinarians (88/113) systematically use the intraperitoneal injection route, which is not registered. Intramuscular injection and injection between muscle layers during surgery are carried out by 82/113 and 43/113 respondents, respectively. Twenty-two respondents apply intra-uterine antibiotics. Most veterinarians (87/113) combine several administration routes. The dosage of antibiotics varies enormously and excessive injection volumes are common, especially when multiple injection routes are combined. Our results show a striking lack of consistency in the antibiotic therapy during elective CS by rural veterinarians. Whereas the drug of choice and the treatment duration were largely in line with the current guidelines, this is certainly not the case for the dosage and the administration route. The intraperitoneal injection of antibiotics cannot be justified. Incorrect dosage of antibiotics should be avoided at all cost. The use of second and third line molecules and the simultaneous use of different antibiotic classes should be discouraged. A major challenge lies in the education of veterinary students and the sensitization of practitioners to avoid or unlearn unnecessary habits concerning antibiotic use. Finally, more clinical trials are needed in order to refine the current guidelines for antibiotic use and to determine the optimal drug of choice, treatment moment and application route in elective caesarean section in cattle.

The association between indicators of inflammation and liver variables during the transition period in high-yielding dairy cows: an observational study.

During the transition period, cows are confronted with infectious and inflammatory challenges leading to an acute phase response (APR) marked by increased hepatic synthesis of positive acute phase reactants (+AP) and a decrease in negative acute phase reactants (-AP). The aim of this study was to quantify the APR in 21 high-yielding dairy cows studied from 9 days before until 42 days after calving, and to assess the association between the APR, disease incidence and indicators of liver function. Repeated blood samples were analyzed for -AP (retinol, albumin, cholesterol), +AP (haptoglobin, caeruloplasmin), paraoxonase, and liver-associated variables (aspartate aminotransferase, γ-glutamyl transferase, bilirubin). All cows displayed postpartum decreases in -AP and paraoxonase, and increases in +AP and liver variables. When retrospectively categorized, cows presenting a stronger -AP decline displayed higher +AP and liver variables, and a higher disease incidence compared to cows with a milder decline. Altogether, typical changes in -AP and +AP identify the transition period as a time of increased inflammatory load. Group differences in liver variables suggest that a more severe APR may be associated with altered liver function. However, no causal relationship can be proven based on this observational dataset, and results should be interpreted cautiously.

Teaching transrectal palpation of the internal genital organs in cattle.

In this article, a simulation model for rectal palpation teaching in cows, Breed'n Betsy, is evaluated. Furthermore, the learning process of rectal palpation is depicted during a training period in live cows. In experiment 1, eight students were trained in live cows (group A) and nine students were trained using Breed'n Betsy (group B). After 25 palpations, their ability to localize and evaluate structures was evaluated in practical tests in live cows. Group A had higher results than group B (p<0.001) and were more skilled at localizing the uterus and localizing and evaluating the ovaries (p<0.05). Group B was better at pregnancy diagnosis (nonsignificant). Results suggest that Breed'n Betsy cannot fully replace training in live cows, but may be a valuable addition to the classical teaching method. Suggestions for future improvement are made. In experiment 2, 10 students were intensely trained in live cows throughout the year and evaluated in practical tests at three time points (September, January, and March). Results were analyzed as a function of time point and the category of experience (1: 0-50 cows; 2: 50-100 cows; 3: 100-150 cows; 4: 150-200 cows; 5: >200 cows). Results increased in time (p<0.05) and were higher in categories 3, 4, and 5 than in category 1 (p<0.05). Although all of the students in the higher categories successfully localized the cervix, uterus, and ovaries, they had difficulties in interpreting these structures, suggesting that palpation of 200 cows is insufficient to reach a consistent level of expertise.