Bacteriological Culture Results Research Articles

Case of delayed diagnosis of combined urogenita and pulmonary tuberculosis in a patient with abnormal uterine bleeding

According to the World Health Organization tuberculosis remains a global health issue for both adult and pediatric populations worldwide. Among women the peak incidence occurs during the active reproductive age, with pregnant women being 1.5–2 times more likely to be affected and experience more severe symptoms than non-pregnant women. In the Altai Region, the epidemiological situation of tuberculosis is characterized by instability, with the incidence rate among women reaching30.5 per 100 000 of the corresponding population in 2022. Delayed diagnosis and treatment can lead to serious complications, including fatal outcomes for both mother and fetus. This article describes a clinical observation of a case of urogenital and pulmonary tuberculosis diagnosed three months after premature abdominal delivery. The course of pregnancy was complicated by acute pleuritis at 13–14 weeks, however, negative results of bacteriological culture of pleural fluid led to the absence of specific tuberculosis treatment and the development of chronic subcompensated placental dysfunction. Three months after delivery, a 34-year-old patient was admitted to the gynecological department of the emergency hospital with abnormal uterine bleeding. The diagnosis was confirmed by histopathology of an endometrial sample revealing tuberculous granulomas without caseous necrosis with epithelioid giant cells of Pirogov – Langhans. Subsequently, generalized tuberculosis was diagnosed in the preterm infant. The negative impact of undiagnosed combined urogenital and pulmonary tuberculosis on the course and outcome of pregnancy is demonstrated, including the development of anemia, placental dysfunction, preterm delivery, puerperal complications, and adverse perinatal outcomes.

Comparative Profiling of Milk Somatic Cells Proteomes Revealed Key Players in Mammary Immune Mechanisms During Mastitis in Tropical Sahiwal (Bos indicus) Cows.

Bovine mastitis poses a significant economic burden on the dairy industry worldwide. This pioneering proteomic study conducted a comparative profiling of milk somatic cell (SC) proteins contributing to mammary immune defense during subclinical and clinical mastitis (CM) in Sahiwal (Bos indicus) cows. Based on California mastitis test (CMT) scores, milk SC counts, differential leukocyte counts (DLCs), and bacteriological culture results, quarter milk SC samples were categorized into healthy (H), subclinical mastitis (SCM), and CM groups. Comparative proteome profiling of milk SCs was done using a label-free liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) proteomic approach. The identified upregulated proteins in mastitis groups such as Vanin 2, Thioredoxin reductase-like selenoprotein T, Ceramidase, Lymphocyte antigen 75, Misshapen-like kinase 1 (MINK1), Thrombospondin 1, Macrophage scavenger receptor 1, Leupaxin, and Lipoamide acyltransferase, involved in immune responses. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed immune functions and pathways like antigen processing, complement cascades, extracellular matrix receptor interaction, efferocytosis, leukocyte migration, chemokine, peroxisome proliferator-activated receptors (PPARs), and transforming growth factor (TGF)-beta signaling. These findings provide essential information on proteomic profiling in milk SCs and contribute valuable insights into immune-related proteins regulated during mastitis in dairy cows. Further, validated proteins (Vanin 2, MINK1, and Thrombospondin 1) offer potential inflammatory biomarkers for early mastitis detection in dairy cows.

Patterns of Microbial Infections in Post-Infective Hydrocephalus and Treatment Methods at Bugando Medical Center

Background: Post Infective Hydrocephalus is among the complicated types of hydrocephalus and poses a challenge in management of these patients, globally. Antibiotics susceptibility patterns of common bacteria implicating in pyogenic ventriculitis complicated with hydrocephalus were not clearly known at our setting. Therefore, we determined antibiotics susceptibility patterns of common bacteria implicating in pyogenic ventriculitis complicated with hydrocephalus to recommend empirical therapy while waiting for culture and sensitivity results at Bugando Medical Centre (BMC) in Mwanza, Tanzania. Methods: Thirty-five patients with hydrocephalus who had undergone cerebrospinal fluid (CSF) tapping and analysis were selected randomly retrospectively and their bacteriological culture and sensitivity results were analyzed to determine the common microbial patterns. Patients were rationally prescribed antibiotics depending on laboratory results for 7 days and were later done control CSF cultures until negative cultures were achieved. Results: A total of 35 patients with median (interquartile range [IQR]) age was 5.5 [2-8] months were reviewed. The majority of participants were males, 62.9% (22/35). 20% (7/35) and 37.1% (13/35) had turbid CSF-shunt sample and first culture positive results, respectively, whereas one patient had culture positive with two pathogens resulting to 14 isolates. Gram positive bacteria were predominantly isolated (66.7%, 8/14), whereby S. aureus was frequently encountered (35.7%, 5/14). Overall, gram positive and negative bacteria exhibited least resistance against tetracycline (16.7% and 25%) and ciprofloxacin (16.7% and 25%) respectively. Three (8.6%) patients died from respiratory complications. Conclusion: Post infective hydrocephalus is a clinically challenging but treatable form of hydrocephalus and Staphylococcus aurous is the most common cause. This study found that the use of a seven-day regimen as cost effective and associated with a shorter hospital stay with the focus on CSF analysis with subsequent microbial targeted antibiotic therapy to achieve sterile CSF culture. Treatment strategy should be multidisciplinary approach with collaboration from microbiologists, radiologists, pediatricians and neurosurgeons.

Laboratory evaluation of a rapid diagnostic test for dairy mastitis.

Rapid diagnostic tests that differentiate between Gram positive, Gram negative and the absence of aerobic bacteria in milk samples from dairy cows with clinical mastitis can support antimicrobial treatment decisions and contribute to a more prudent use of antimicrobials in the dairy industry. The objective of this study was to evaluate the test characteristics of the novel rapid BACT mastitis test in discriminating causes of clinical mastitis under laboratory conditions. Test outcomes of 155 milk samples from clinical mastitis cases were incubated for 14-16 h in the BACT test and compared to results of bacteriological culture. The accuracy for detection of bacterial growth and Gram positive growth was 91 and 89%, respectively. The BACT test could provide an accurate and relatively fast decision tool for farmers to aid in antimicrobial treatment decisions in cases of clinical mastitis.

RESULTS OF BACTERIOLOGICAL MONITORING OF URINARY TRACT INFECTIONS IN CHILDREN OF ZAPORIZHZHIA REGION IN 2023

The aim of the study. To investigate the structure of pathogens of acute urinary tract infections in children of Zaporizhzhia region in 2023 and their sensitivity to antibacterial agents. Materials and methods. The analysis of the structure of pathogens in acute urinary tract infections and the determination of their sensitivity to antibiotics based on the results of bacteriological urine cultures of 322 children with acute urinary tract infections treated at the Zaporizhzhia Regional Children's Hospital in 2023 was performed. For each isolate, the multidrug resistance index was calculated, which allowed to judge the relative integral resistance of the isolated microorganisms to antibacterial drugs. Results. It was established that the most common cause of urinary tract infections in children were gram-negative microorganisms (61.0%), including Escherichia coli (37.4%) and Klebsiella pneumoniae (10.8%) which were most often verified. Gram-positive bacteria were identified in 38.5% of patients, among which the Enterococcus bacteria (29.2%) predominated. High resistance of Escherichia coli to amoxicillin/clavulanate (31.5%), ciprofloxacin (30.1%), cefepime (20.1%), and ceftazidine (20.5%) was detected, including 29 (39.7%) multidrug-resistant isolates of Escherichia coli. The highest resistance of Klebsiella pneumoniae was found to amoxicillin/clavulanate (76.2%), cefepime (33.3%), ceftazidine (33.3%), ciprofloxacin (42.8%), and phosphamycin (28.6%). Multidrug resistance was shown by 76.2% of Klebsiella pneumoniae isolates. The highest susceptibility of Enterococcus bacteria was found to ampicillin (98.2%), linezolid (98.2%), vancomycin (100%), nitrofurantoin (100%), and tigecycline (100%). In no case microorganisms showed complete sensitivity to imipenem, and 17.9% of Enterococcus isolates were resistant to this antibiotic. Conclusions: At the present stage, the dominance of gram-negative microflora among the pathogens of urinary system infections in children remains, among which Escherichia coli (37.4%) continues to be the leading pathogen. The growing role of Enterococcus faecalis (29.2%) as an etiologic factor of urinary tract infections in children was noted, which should be taken into account during the empirical use of antibiotics, especially if there is no effect within the first two days after the start of therapy.

The coexistence of several microbial species at the same site of bovine mammary gland parenchyma infection and their mixed infections

<b>The purpose of this study was to analyse both negative results of bacteriological cultures and the coexistence of several pathogens at the same intramammary inflammatory focus within bovine udder parenchyma in order to compare the health status of cows’ udders in 1985 and 2021. Mastitis was diagnosed on the basis of anamnesis, clinical inspection of the udder, milk quality, and somatic cell counts (SCC on the Fossomatic 5000-FC). In 1985, joint bacteriological and mycological diagnosis was based on the guidelines established for bovine mastitis by FIL-IDF (1971), while for 2021, it was based on the National Mastitis Council (NMC) laboratory Handbook on bovine mastitis (Hogan et al., 1999). In the first study (1985), mainly the genus Staphylococcus was isolated, but at present other species may be involved in cases of mastitis. In both periods, both subclinical and clinical cases were registered. In addition, the milk yield of cows was observed to increase over time, with concomitant deterioration of mammary gland immunity. The phenomenon of biofilms was documented in the study, which may be explained not only by the low efficacy of antibiotic therapy against udder disease but also by impaired immunity in cows, as the lymphocytes trapped in the biofilm matrix were inactive with respect to both cellular and humoral responses.

The Extent Response of Bacteria Isolated from Cases of Acute, Subacute and Chronic Mastitis in Iraqi Camels to some Antibiotics used in Treatment

Our study was carried out on (75) dairy camels, aged (7-22) years, from Iraqi camels for each of the governorates (Babylon-Karbala-Al-Qadisiyah) with (25) camels from each governorate for the time period 1/6/2021 until 1/12/2022 to isolate and diagnosis the bacteria causing acute, subacute and chronic mastitis in camels, examining their sensitivity to common antibiotics used in treatment. The results of the study showed that the number of camels infected with mastitis for all types was (51) camels with a rate of (68%) out of a total of (75) milking camels, and the number of healthy camels was (24) with a rate of (32%). The number and percentage of subacute mastitis was (26) with a rate of (35%), acute mastitis (10) with a rate of (13%), and chronic mastitis (15) with a rate of (20%). As for the results of mastitis on the basis of inflammation tests for each quarter of the udder, the highest rate was in two quarters, where the percentage of infected camels was (35%), while the lowest percentage of infection was in four quarters or the whole udder with a rate of (18%). Results of bacteriological culture of milk samples of camels infected with mastitis, which numbered (306) samples. The highest percentage of isolates recorded was of C.N. Staphylococci by (35.62%), while the percentage of Staph.sciuri, Staphylococcus aureus and Staph.epidermidies respectively was (16.66%), (13) and (10.13%). The percentage of Streptococcus agalactia, Pasteurella haemolytica, Arcanobacterium pyogenes and E. coli respectively (10.45%), (6.2%), (5%) and (2.94%). The results of the sensitivity and antibiotic resistance examination, we show that there is statistical significant difference at the level of p ≥ 0.05 between the groups of the ages of camels.

Analysis of etiologic structure and antibiotic resistance of the main pathogens of infectious complications in patients with acute leukemia

Objective. Analyze the etiological structure of infectious complications in patients with acute leukemia and determine the sensitivity to antibacterial drugs of the main pathogens.Materials and methods. The results of bacteriological cultures and antibiograms of 177 patients with acute leukemia and infectious complications for the period 2019–2021 were retrospectively studied.Results. In 2019, the frequency of isolation of Gr+ flora was 45.28% and Gr- was 54.72%; in 2020 the percentage of Gr+ bacteria was 34.62%, Gr− was characterized by 65.38%; in 2021 it was 41.3% and 58.7% respectively, (x2=0.20, p=0.66). In 2019, one of the main pathogens was Klebsiella pneumoniae (21%), which retained its positions by 2021 (13%; x2=0.05, p=0.82). The causative agent showed polyresistance to many antibiotics, except for imipenem (100%) and fosfоmycin (100%). In the period 2020–2021 Klebsiella pneumoniae began to compete with Stenotrophomonas maltophilia, which showed an increase in the detection rate from 11% to 17% (x2=29.46, p=0.0001). Having polyresistance, the pathogen showed 100% sensitivity only to ticarcillin/clavulanate and co-trimoxazole.Conclusion. In the absence of changes between the frequency of isolation of Gram-positive and Gram-negative microorganisms, the increase in the number of strains of Gram− microorganisms (K. pneumoniae and S. maltophilia) with multiple antibiotic resistance among the causative agents of infectious complications in patients with acute leukemia necessitates continuous microbiological monitoring to improve the tactics of antibacterial drugs.

Determination of microbial contamination on the outer surface of needleless connectors before and after disinfection

ObjectivesThe aim of this study was to determine bacterial contamination of needleless connectors before and after disinfection to assess the risk for the point of catheter-related bloodstream infections. Research methodology/designExperimental study design. SettingThe research was carried out on patients with a central venous catheter hospitalized in the intensive care unit. Main outcomes measuresBacterial contamination of needleless connectors integrated into central venous catheters was assessed before and after disinfection. Antimicrobial susceptibilities of colonized isolates were investigated. In addition, the compatibility of the isolates with the bacteriological cultures of the patients was determined over a one-month period. ResultsBacterial contamination varied between 5×103 and 1×105 colony forming unit was detected before disinfection in 91.7% of needleless connectors. Most common bacteria were coagulase-negative staphylococci; others were Staphylococcus aureus, Enterococcus faecalis, and Corynebacterium species. While most isolates were resistant to penicillin, trimethoprim-sulfamethoxazole, cefoxitin, and linezolid, each was susceptible to vancomycin or teicoplanin. Bacterial survival was not detected on needleless connectors after disinfection. There was no compatibility between the one-month bacteriological culture results of the patients and the bacteria isolated from the needleless connectors. ConclusionBacterial contamination was detected on the needleless connectors before disinfection, although they were not rich in bacterial diversity. There was no bacterial growth after disinfection with an alcohol-impregnated swab. Implications for clinical practiceThe majority of needleless connectors had bacterial contamination before disinfection. Needleless connectors should be disinfected for 30 seconds before use, particularly in immunocompromised patients. However, the use of needleless connectors with antiseptic barrier caps may be a more practical and effective solution instead.

The effect of antibiotic versus no treatment at dry-off on udder health and milk yield in subsequent lactation: A retrospective analysis of Austrian health recording data from dairy herds.

Bovine mastitis is the most commonly diagnosed disease of dairy cows worldwide and causes extensive economic losses to milk producers. Intramammary infection status before dry-off plays a decisive role with respect to udder health and milk yield in the subsequent lactation. The aim of this study was to compare the effect of antibiotic dry cow therapy (DCT) versus no treatment at dry-off on milk yield, somatic cell count (SCC), inflammation of the mammary gland (IMG), and the incidence of clinical mastitis in the subsequent lactation. Dairy herd data from 251 Austrian dairy farms were recorded over an observation period of 12 mo and subsequently analyzed. The data set included 5,018 dairy cows: 2,078 were treated with antibiotics (abDCT group) and 2,940 were not treated (noDCT group) at dry-off. The abDCT group was subdivided, based on the antimicrobial active substances used for drying off, into 4 different groups (penicillins, cloxacillin, cephalosporins, and rifaximin). Based on bacteriological culture results, infections were grouped into those caused by major, minor, and other pathogens. Additionally, the IMG was defined via SCC from milk recording data using a cutoff of 200,000 cells/mL before drying off and after calving. The incidence of clinical mastitis cases within 30 and 90 d in milk was calculated using veterinary diagnosis data. To investigate the effect of different dry cow therapies on the following parameters: milk yield, SCC, and diagnosed clinical mastitis cases, different linear mixed models were constructed. Overall, the abDCT group was determined to have a significantly higher milk yield over 305 d in milk in the subsequent lactation (increase of 6.18%), compared with the noDCT group (increase of 4.29%). Both groups (abDCT and noDCT) demonstrated a decrease in the first SCC after calving compared with the SCC before dry-off, although the treated cows had a significantly higher reduction. Regarding the different antibiotic groups, with exception of the rifaximin treated cows, all antibiotic groups showed a significant difference from not treated cows with respect to SCC. Additionally, we were able to demonstrate that cows with IMG before dry-off had a 2.073 times higher chance of an increased SCC (>200,000 cells/mL) after calving. With respect to the veterinary diagnosis data, neither the IMG before drying off nor the type of DCT had a significant influence on the probability of developing clinical mastitis within 30 or 90 d in milk. Only a small number of treatments was accompanied with a bacteriological examination before drying off. However, the existing data in this study indicates that the intramammary infection status before dry-off in combination with different dry cow treatments influences udder health and milk yield after calving. Nevertheless, further studies with larger data sets of bacteriological examinations are necessary to enable a more in-depth investigation into the effects of different antibiotic substances used for DCT.

ПРИНЦИПЫ ПРОФИЛАКТИРОВАНИЯ АНТИБИОТИКОРЕЗИСТЕНТНОСТИ ПРИ ЛЕЧЕНИИ УРОЛОГИЧЕСКОГО СИНДРОМА КОШЕК

The article presents data on the occurrence of urological syndrome in cats of different age and gender groups and the structure of diseases of the lower urinary tract, leading to its development. The most frequently detected pathology was idiopathic feline cystitis (IFC) 82.3 %, urocystolithiasis and bacterial cystitis were less frequently detected – 10.3 % and 7.4 %, respectively. In the differential diagnosis of recurrent cystitis by bacteriological seeding of urine, it was found that more often the cause of the resumption of dysuria was IFC – 73.7 % of negative urine seeding. In other cases, the bacterial etiology of bladder inflammation was confirmed and uropathogens were identified: E. faecalis, E. faecium – 10.5 %, P. mirabilis – 5.3 % of results. When diagnosing cystitis in cats, the following algorithm should be followed: the primary instrumental diagnostic method is ultrasound (the fact of cystitis is established: thickening of the bladder wall, changes in its structure, the presence of suspension and / or concretions in the organ cavity), to differentiate the etiology of cystitis, it is necessary to use laboratory diagnostic methods – general analysis and bacteriological urine culture. Drug treatment consisted of anesthesia (idiopathic cystitis), and in case of septic cystitis, etiotropic therapy was additionally prescribed. Analgesia was carried out in a complex with the use of drugs from the group of nonsteroidal anti-inflammatory drugs (NSAIDs) – meloxicam 0.05 mg/kg 1 time per day and alpha-blockers – terazosin as an antispasmodic at a dose of 0.5 mg /cat 2 times a day. Antibacterial therapy was used strictly according to the results of bacteriological urine culture: the first–line drug of choice was used – amoxicillin with clavulanic acid, reserve antibiotics – cefixime, marbofloxacin; dosages, frequency of administration per day, the course of treatment was prescribed to each patient individually, according to the diagnostic results. The algorithm of actions of a veterinarian when working with cystitis in cats, developed during this study, will help to avoid uncontrolled use of antibiotics, and, as a consequence, the development of antibiotic resistance

Evaluation of a droplet digital PCR assay for quantification of Mycobacterium avium subsp. paratuberculosis DNA in whole-blood and fecal samples from MAP-infected Holstein cattle.

Bovine paratuberculosis (PTB) is an infectious disease that affects ruminants worldwide and is a burden on the dairy industry. PTB control measures include culling of Mycobacterium avium subsp. paratuberculosis (MAP)-infected animals from the herd and the enhancement of farm-biosecurity measures. Diagnostics tools for the direct detection of MAP are fecal real-time qPCR and bacteriological culture, the last one being considered the gold standard. However, both show limitations for detecting subclinical MAP-infected cattle with low bacterial load in feces and gut tissues. Droplet digital polymerase chain reaction (ddPCR) is a third-generation PCR method that shows high reproducibility for the quantification of low DNA copy numbers. The objective of this study was to design a ddPCR assay to detect and quantify a fragment of the F57 MAP-specific sequence in samples of naturally MAP-infected Holstein cattle. DNA was isolated from whole-blood and fecal samples from control cows with a negative ELISA and qPCR result (N = 75) and from cows with PTB-associated focal (N = 32), multifocal (N = 21), and diffuse lesions (N = 17) in gut tissues. After ddPCR, the DNA extracted from fecal samples of cows with diffuse lesions showed higher mean copies per microliter (13,791.2 copies/μl) than samples from cows with multifocal lesions (78.8 copies/μl), focal lesions (177.1 copies/μl) or control cows (4.8 copies/μl) (P ≤ 0.05). Significant differences in mean DNA copies/μl were also observed in the blood samples from cows with focal lesions (47.7 copies/μl) when compared with cows with multifocal and diffuse lesions; 18.1 and 12.4 copies/μl, respectively. Using a principal component analysis, the results of the fecal ddPCR clustered together with the results of a commercial ELISA for the specific detection of MAP antibodies, fecal and tissue qPCR, and bacteriological culture results. In contrast, blood ddPCR results clustered together with the results of an ELISA for the detection of a biomarker of subclinical PTB, the ABCA13 transporter. Blood ddPCR was the most sensitive tool (sensitivity 71%, specificity 100%) of all the quantitative methods used in the study for the detection of subclinical cows with focal lesions.

Epidemiology and Early Bacteriology of Extremely Severe Burns from an LPG Tanker Explosion in Eastern China

The incidence of liquefied petroleum gas (LPG)-related accidents in China has increased over the recent years. In addition, infection remains a big challenge in cases of severe burns. Therefore, the present study aimed to provide valuable information for a better control of infections in the event of such disasters. In this study, a total of 16 patients who suffered extremely severe burns after an LPG tanker explosion were included. Thereafter, bacteriological culture results were collected within a week. Of 16 patients, 13 (81.25%) were male and the average age of all patients was 60.63 years. In addition, the mean burned area was 83.03% TBSA. Additionally, a total of 553 organism cultures were conducted out of which 287 isolates (51.90%) showed positive results. Notably, 38.52% were Gram-negative bacteria, 7.59% were Gram-positive bacteria and 5.79% were fungi. Moreover, the most prevalent Gram-negative bacteria were Stenotrophomonas maltophilia (28.97%) followed by Acinetobacter baumannii (28.53%), and Klebsiella pneumoniae (14.02%). On the other hand, the three most predominant Gram-positive bacteria were Enterococcus faecalis (33.33%), Staphylococcus aureus (28.89%) and Staphylococcus sciuri (17.78%). Furthermore, the most common fungi included Candida (38.24%), Fusarium (20.59%) and Aspergillus fumigatus (14.71%). With regard to the bacterial resistance patterns, carbapenem-resistant organisms included Acinetobacter baumannii (97.80%), Pseudomonas aeruginosa (67.57%), and Klebsiella pneumoniae (75.56%). In addition, Staphylococcus sciuri, Staphylococcus epidermidis, and Staphylococcus haemolyticus were identified to be methicillin-resistant. This study revealed that there was a high incidence of infection in victims of severe burns as a result of mass burn accidents, accompanied by early fungal infection.

Microscopical Evaluation of Smears of the Leptomeninges to Predict Meningitis in Piglets.

Simple SummaryMeningitis (inflammation of the membranes covering the brain) is a common problem in 3- to 10-week-old piglets. It is often caused by bacteria that are called Streptococcus. Clinical signs such as problems with coordination of the limbs can be observed by the farmer, but it is also possible that piglets die suddenly, before clinical signs are detected. Therefore, examining dead piglets is often performed to establish a diagnosis. Unfortunately, meningitis is often difficult to recognise by the eye. To assist veterinarians in forming a quick diagnosis of meningitis and possible bacterial involvement, a microscopic evaluation of cells taken from the meninges was performed. This examination allowed the prediction of the likely presence of meningitis and the involvement of Streptococcus species in most of the cases. Microscopic findings were compared with bacteriological culture results, and based on this comparison, indications of meningitis and Streptococcal presence seemed correct in 89% and 100% of the cases, respectively. However, if only a few cells indicating meningitis were observed microscopically, no reliable prediction was possible. In conclusion, microscopic evaluation of cells from the meninges of piglets can help swine practitioners in establishing a tentative diagnosis of meningitis more quickly, which allows for better treatment, improves animal welfare, and reduces antimicrobial use.Meningitis, caused by bacterial infections such as Streptococcus spp., is a frequently observed cause of death in pigs. In order to implement effective treatment and avoid further losses, it is important to establish this diagnosis quickly. However, this is often difficult because macroscopic lesions may not be visible, and additional laboratory testing may take several days. The present study investigated whether microscopical examination of impression smears of the leptomeninges taken during necropsy can help in establishing a presumptive diagnosis of meningitis in pigs more quickly. In total, 54 pigs suffering from neurological signs and/or (acute) mortality were examined. They were 3 to 10 weeks old and originated from 16 farms. From each pig, bacterial culture was performed on one half of the brain using a swab from the leptomeninges. From the other half, paired cytological impression smears of the leptomeninges were stained with a commercial quick stain dye (Hemacolor®) and Gram stain and microscopically evaluated for the abundance of neutrophils and the presence of short-chain coccoid bacteria. Bacterial culture of the leptomeninges was positive in 36/54 cases, in 28 of which Streptococcus spp. were found. The numbers of smears with low, moderate, or high abundance of neutrophils were 19, 17, and 18, respectively. Short-chain coccoid bacteria were detected successfully in 12 pigs in the Gram-stained smear. The positive predictive value of smears with moderate or high abundance of neutrophils for bacterial presence and, therefore, likely meningitis was 89%, whereas the negative predictive value of smears with low abundance of neutrophils was 74%. The positive predictive value of smears with short chains of coccoid bacteria for diagnosis of Streptococcus spp. was 100%, whereas the negative predictive value was 62%. In conclusion, microscopical examination of impression smears of the leptomeninges of piglets with neurological signs and/or (acute) mortality is a feasible procedure that can help swine practitioners in establishing a tentative diagnosis of meningitis more quickly, especially if neutrophils are abundant, and short chains of coccoid bacteria are present.

Microflora of sputum and autopsy material of patients with COVID-19.

The rapid spread of a new coronavirus infection in the country actualizes the conduct of bacteriological studies of clinical material obtained from the respiratory tract of patients with COVID-19. During the experiments, 230 sputum samples and 260 autopsy lung samples from patients with COVID-19 were analyzed. 946 high-risk strains were isolated and identified by MALDI-ToF mass spectrometry on a Microflex LT instrument (Bruker®). According to the results of bacteriological cultures of sputum, a predominance of gram-positive ones was revealed, amounting to 50.5% (222 strains) of the total number of isolated pathogens. However, falling into this group is manifested by natural representatives of the microflora of the human mucous membranes from the genera Streptococcus, Rothia and Lactobacillus (109 strains in total), which can be manifested by the detection of improper sputum collection, causing contamination by the substance of intense salivation and nasopharyngeal discharge. In turn, the "classic" gram-positive causative agents of pneumonia were detected much less frequently: S. aureus in 5 cases, S. pneumoniae in 6 patients. The causative agents in the order Enterobacterales are represented by 42 strains, among which the most likely species are K.pneumoniae (27 strains). In the group of non-fermenting gram-negative bacteria, A. baumanii (29 strains) prevailed, and P. aeruginosa was also identified in 2 cases. When analyzing the results of a microbiological study of autopsy material (lungs) of patients with COVID-19, significant differences in the qualitative and quantitative composition of the microflora were revealed, compared with sputum. In the group of gram-positive bacteria, 15 strains of the natural microflora of the mucous membranes were identified, while sensitive species dominated among gram-negative pathogens: K. pneumoniae (102 strains), A. baumanii (75 strains), P. aeruginosa (11 strains). Regular microbiological monitoring is essential for antibiotic therapy and prevention of secondary bacterial infection. In the event of a fatal outcome, the results of microbiological analysis of autopsy material can determine the cause of death of the patient.

A Genome-Wide Association Study for Tolerance to Paratuberculosis Identifies Candidate Genes Involved in DNA Packaging, DNA Damage Repair, Innate Immunity, and Pathogen Persistence

Although the genetic susceptibility to diseases has been extensively studied, the genetic loci and the primary molecular and cellular mechanisms that control disease tolerance are still largely unknown. Bovine paratuberculosis (PTB) is an enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP). PTB affects cattle worldwide and represents a major issue on animal health. In this study, the associations between host genetic and PTB tolerance were investigated using the genotypes from 277 Spanish Holstein cows with two distinct phenotypes: cases) infected animals with positive PCR and bacteriological culture results but without lesions in gut tissues (N= 24), and controls) animals with negative PCR and culture results but with PTB-associated lesions (N= 253). DNA from peripheral blood of the study population was genotyped with the Bovine EuroG MD Bead Chip, and the corresponding genotypes were imputed to whole-genome sequencing (WGS) data. A genome-wide association study was performed using the WGS data and the defined phenotypes in a case-control approach. A total of 142 single nucleotide polymorphisms (SNPs) were associated (false discovery rate ≤ 0.05, P values between 1.5 × 10-7 and 5.7 × 10-7) with tolerance (heritability= 0.55). The 40 SNPs with P-values < 5 × 10-7 defined 9 QTLs and 98 candidate genes located on BTA4, BTA9, BTA16, BTA25, and BTA26. Some of the QTLs identified in this study overlap with QTLs previously associated with PTB, bovine tuberculosis, mastitis, somatic cell score, bovine diarrhea virus persistent infection, tick resistance, and length of productive life. Two candidate genes with important roles in DNA damage response (ERCC4 and RMI2) were identified on BTA25. Functional analysis using the 98 candidate genes revealed a significant enrichment of the DNA packaging process (TNP2/PRMI1/PRM2/PRM3). In addition, the TNF-signaling (bta04668; TRAF5/CREB5/CASP7/CHUK) and the toxoplasmosis (bta05145; TGFβ2/CHUK/CIITA/SOCS1) pathways were significantly enriched. Interestingly, the nuclear Factor NF-κβ Inhibitor Kinase Alpha (CHUK), a key molecule in the regulation of the NF-κB pathway, was enriched in both pathways. Taken together, our results define a distinct immunogenetic profile in the PTB-tolerant animals designed to control bacterial growth, modulate inflammation, limit tissue damage and increase repair, thus reducing the severity of the disease.

Pathological features of Brucella spondylitis: A single-center study

ObjectiveTo explore the pathological features of Brucella spondylitis (BS) under the optical microscope, thus providing pathological references for the diagnosis. MethodsWe retrospectively analyzed 70 BS patients (42 males and 28 females, mean age 52.01 ± 10.77 [20–74] years) admitted in the Department of Spine Surgery, the General Hospital of Ningxia Medical University, from January 2013 to December 2020. Their medical history, clinical manifestations, laboratory test results, imaging findings and bacteriological culture results were collected. Among them, 5, 5, 43, 4 and 13 cases demonstrated involvement into the cervical vertebra, thoracic vertebra, lumbar vertebra, thoracolumbar vertebra and lumbosacral vertebra, respectively. Notably, L4 showed pathology in 32 cases. Pathological features of BS were analyzed by H&E staining of granulation tissues, sclerotic bones, sequestrums, and intervertebral discs. Results42 cases underwent bacterial culture, of which 4 were positive, and the positive rate of bacterial culture was only 9.5%. The highest Vas score was 7, the lowest was 4, and the average was 5.76 ± 0.89. The highest CRP was 153 mg/L, the lowest was 0.98 mg/L, and the average was 30.98 ± 33.79 mg/L. The highest ESR is 112 mm/h, the lowest is 5 mm/h, and the average is 49.34 ± 27.73 mm/h. Under the optical microscope, BS manifested acute or chronic inflammation. Acute inflammatory features included neutrophil and eosinophil infiltration, necrosis, and abscesses, while chronic inflammatory features included lymphocyte, plasma cell, fibrous tissue and monocyte infiltration, hyaline degeneration, angiogenesis and hyperplasia of other tissues. Other features included vasodilation, hemorrhage, granulomas and multinucleated giant cell infiltration. In the present study, chronic inflammation was observed in 25 cases, in-acute-phase chronic inflammation in 45 cases, and acute inflammation in no cases. Pathological features of BS under the microscope included foam cell reaction in 46 cases, histiocytic reaction in 24 cases and eosinophilic abscesses in 6 cases. Eosinophil infiltration was observed in 45 cases (mainly during the acute phase of chronic inflammation) and massive eosinophilic abscesses in 6 cases. Granulation tissue hyperplasia followed inflammatory repair in 25 BS cases, and was generally boosted in the acute phase of chronic inflammation. Multinucleated giant cell infiltration and granulomas were less observed in BS cases, which differed from pathological features of spinal tuberculosis. ConclusionsChronic inflammation or in-acute-phase chronic inflammation is the main pathological feature of BS, while the single acute inflammation is less observed in BS cases. Foam cell reaction and histiocytic reaction scale up during the acute phase of chronic inflammation, and some BS patients may develop massive eosinophilic abscesses. Granulation tissue hyperplasia, rather than multinucleated giant cell infiltration and granulomas, serve as pathological reference for the differential diagnosis of BS and spinal tuberculosis.

A potential treatment approach for subclinical mastitis in dairy cows: auriculotherapy of the auricular branch of the vagus nerve.

This research aims to test the hypothesis that neural therapy (NT) on the auricular branch of the vagus nerve (ABVN) in dairy cows diagnosed with subclinical mastitis (SCM) results in a reduction of the somatic cell count. Therapeutic options for SCM are mostly based on use of antibiotics and often lead to unsatisfactory results. An alternative therapy targeting the anti-inflammatory properties of the vagus nerve showing good efficacy, economic viability and without major side effects would be of considerable interest. Auriculotherapy (AT) was performed using three repeated infiltrations of 8.0 mg (0.4 ml) procaine hydrochloride (2%) at the location of the ear tag associated with the auricular acupuncture point (AAP) of the udder. Some 85 clinically healthy cows from nine dairy farms were sampled for evaluation of quarter somatic cell count (QSCC) on four days (d0, d2, d4 and d6). Quarters with a QSCC > 100 000 cells/ml on d0 were included in the analysis. Over the study period, a total of 784 quarters were analysed, 385 control (CON) quarters from 40 cows and 399 treated (TRE) quarters from 45 cows. Results showed that AT of the ABNV resulted in a significant reduction of the QSCC after three treatments. The effect was independent of bacteriological culture results of the quarter milk samples. The bacteriological cure rate, however, was not influenced by AT. To our knowledge this is the first report of AT reducing QSCC in dairy cows with SCM. Before AT can be regarded as an alternative therapeutic approach, further research should focus on possible long-term effects of AT on the reduction of SCC, any bacterial elimination and the neural pathways of AT in dairy cows with SCM.

Correlation of Direct Smear Observation and Bacteriologic Culture Results in a Tertiary Care Hospital

Correlation of Direct Smear Observation and Bacteriologic Culture Results in a Tertiary Care Hospital

Using the Idexx SediVue Dx to predict the need for urine bacteriologic culture in cats.

We analyzed urine samples from 191 cats for bacteriuria with an automated urine sediment analyzer (Idexx SediVue Dx), combined with image review by an observer, and compared to bacteriologic culture results. Sixty-nine samples were unambiguously assigned to be free of bacteria by the instrument and the observer, and no bacterial growth was detected. Twenty-seven samples were unambiguously assigned to have bacteriuria; 24 of these 27 samples were culture-positive. For these samples, bacteriuria was predicted with a sensitivity of 100% and a specificity of 96%. A clear assignment was not possible for 95 samples, 81 of which were culture-negative. Specificity dropped to 45% when all samples were considered. Using the automated leukocyte count to predict bacteriuria, sensitivity was 82% and specificity was 75%. Automated sediment analysis is faster and less observer-dependent than sediment analysis under a microscope, but accurate detection of bacteriuria remains difficult in a large proportion of samples. Bacteriuria was significantly associated with leukocyte count; the leukocyte count was >5/high power field in 82% of culture-positive samples.