Sheep Blood Research Articles

Improving the microbiological diagnosis of fracture-related infection and prosthetic joint infection through culturing sonication fluid in Bactec blood culture bottles.

Sonication of surgically removed implants appears to optimize the microbiological diagnosis in orthopedic implant-associated infections (OIAI). However, reports of infection with negative cultures can still reach high rates. A study evaluating the inoculation of sonication fluid into blood culture bottles (SFBCB) in patients with fracture-related infection (FRI) and prosthetic joint infection (PJI) is necessary. This study compared the accuracy SFBCB over the conventional sonication fluid cultures (CSFC) and tissue culture (TC). Consecutive patients who underwent implant removal surgeries due to any reason had their implants sonicated according to standardized method. Definitions of PJI and FRI were based upon criteria by European Bone and Joint Infection Society (EBJIS) and Metsemakers, respectively. Between three to five intraoperative tissue samples were processed. The implant`s sonication fluid was seeded onto sheep blood agar, chocolate agar, thioglycolate broth and on tryptic soy broth for CSFC, while was also inoculated into blood culture bottles and incubated in the automated system during 5 days for SFBCB. Overall, 74 patients were analyzed, of which 57 with OIAI (48 FRI and 09 PJI) and 17 aseptic failures (03 arthroplasties and 14 osteosynthesis). Interestingly, SFBCB demonstrated significantly higher sensitivity compared to CSFC (96.5% [95% CI, 88-100] vs. 78.9% [95% CI, 66-89], p = 0.004), and to TC (96.5% [95% CI, 88-100], vs. 57.9% [95% CI, 44-71], p < 0.001), whereas there were no significant differences in specificity between the three methods. In comparison to CSFC and TC, SFBCB improved sensitivity for diagnosing OIAI without compromising specificity.

Lactococcus garvieae as a Novel Pathogen in Cultured Pufferfish (Takifugu obscurus) in China

In October 2023, a disease outbreak in pufferfish (Takifugu obscurus) farms in Zhongshan City, Guangdong, China, caused high mortality. Diseased fish (mean length: 15 ± 1 cm) exhibited swimming disorders, fin rot, hemorrhage, and an enlarged spleen. Histopathological observations generally revealed inflammation, necrosis, and congestion in the spleen, kidneys, and brain tissues. The most severe pathological changes included interstitial edema and tubular atrophy in the kidneys, hemosiderin deposition in the spleen, massive red blood cell infiltration, and a decrease in lymphocytes. A single strain of bacteria (Tol-1) was isolated from the diseased pufferfish and identified as a Gram-positive streptococcus strain, exhibiting α-hemolysis on sheep blood agar plates. Through biochemical characterization, 16S rDNA sequencing, morphological analysis, and specific primer-based identification, the Tol-1 strain was identified as Lactococcus garvieae, serotype I. Antimicrobial susceptibility testing indicated that Tol-1 was sensitive to Chloramphenicol, Ampicillin, Cephalexin, and Doxycycline, but resistant to Kanamycin, Gentamicin and Ciprofloxacin. In addition, 15 common virulence factors were detected in the Tol-1 strain, including adhPav, adhPsaA, adhC I–II, adh, and hly 1–3. Pufferfish (mean length: 17 ± 1 cm) subjected to artificial infection via intraperitoneal injection (IP) with the Tol-1 strain exhibited clinical symptoms and histopathological damage similar to those observed in naturally infected fish. An infection dose of 1 × 105 CFU/fish resulted in 80% mortality. The study fulfilled Koch’s postulates, indicating that the disease outbreak in pufferfish was caused by L. garvieae, which exhibited a high mortality rate in pufferfish despite the subtle clinical symptoms. These results serve as a warning for pufferfish farming areas and provide a scientific basis for future prevention and control efforts.

B-019 Reduction in Culture Media Costs, without Impacting Turnaround Time, Through Implementation of an Automated Microbiology Platform and Gram Stain Interpretation

Abstract Background Automated plating instruments have been shown to improve quality in Microbiology laboratories in a number of ways including consistent streaking patterns that provide better isolation of bacterial colonies compared to manual methods. The time per culture set up and automated camera imaging is dependent upon the amount of media set, such that more individual plates slow down turn around time. Conversely, smaller media surface area and the use of non-selective media may result in mixed colonies that require sub-culture for isolation before identification and susceptibilities can be performed. Based upon these findings, we attempted to reduce our media usage per culture with the implementation of automation, with the goal of using as few plates as possible while at the same time not negatively impacting the turn around time of the culture via the need to subculture poorly isolated colonies. We conducted a pre-and post-implementation study evaluating the costs per culture and turn around time delays related to subculturing in a subset. Methods In the pre-implementation period, from April-January 2023, three individual media plates (sheep blood agar, chocolate agar and CNA/MacConkey bi-plate) were routinely set for general aerobic bacterial culture sources including swabs and body fluids. In the post implementation period, from April-January 2024, a single sheep blood/chocolate agar bi-plate was set unless the gram stain showed evidence of mixed organisms (defined as more than one morphology) at which time a secondary selective media (CNA/MacConkey) was manually set. For urine cultures, in the pre-implementation period 2 plates were set (sheep blood and CNA/MacConkey bi-plate) and in the post-implementation period a single chromogenic agar/sheep blood bi-plate was set. Total numbers of cultures in both periods were collected as well as the number of cultures for which additional media were set based on gram stain results. Orders of subculture media in the laboratory information system were compared per positive cultures. Statistical analysis was performed in GraphPad Prism. Results In the pre- versus post-implementation periods, a total of 16,454 vs 17,782 general aerobic cultures and 50,680 vs 53,916 urine cultures were set, respectively. Only 3.6% of general cultures required additional media set based on gram stain. The difference in cost between pre- and post-implementation was approximately a two-thirds reduction in cost for general cultures and a 25% reduction for urine cultures, with a total savings of approximately $34,238 in media avoidance in the post-implementation period. The frequency of positive cultures requiring subculture was not significantly different between the pre- (average 36% of positive growth) and post-implementation (average 22% of positive growth) periods for both culture types and may actually have been reduced due to automation. Conclusions The implementation of reduced media concomitant with streaking via automation resulted in significant cost savings while not negatively impacting isolation, which in turn impacts culture turn around times. The automation may have reduced need for subculture, perhaps due to better isolation, thus subsequent analyses will explore this additional finding.

Antimicrobial effects of a multimodal wound matrix against methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa in an in vitro and an in vivo porcine wound model.

Chronic non-healing wounds pose significant challenges due to an elevated inflammatory response caused in part by bacterial contamination (Physiol Rev. 2019;99:665). These wounds lead to billions being spent in the health care system worldwide (N Engl J Med. 2017;376:2367, Int J Pharm. 2014;463:119). We studied the in-vitro and in-vivo antimicrobial effects of a multimodal wound matrix (MWM) against two common wound pathogens, Methicillin-Resistant Staphylococcus aureus (MRSA USA300) and Pseudomonas aeruginosa ATCC 27312 (PA27312) (Int Wound J. 2019;16:634). The in-vitro study conducted was a zone of inhibition test with the two microbes at 104 Log CFU/mL inoculated on Tryptic soy agar with 5% sheep blood (TSAII) plates. Treatments used were MWM, Mupirocin (Positive control for MRSA), Silver Sulfadiazine (Positive Control for PA), Petrolatum and Sterile Saline (both serving as Negative Controls). Treatments were allowed to diffuse into the agar for 3 h and then were incubated for 24 h at 37°C. The in-vivo study utilized a deep dermal porcine wound model (22 × 22 × 3 mm) created on six animals. Three animals were inoculated with MRSA USA300 and the other three with PA27312 with each allowing a 72-h biofilm formation. After 72 h, baseline wounds were assessed for bacterial concentration and all remaining wounds were treated with either MWM alone, Silver Treatment or Untreated Control. Wounds were assessed on days 4, 8 and 12 after treatment application for microbiological analysis. In-vitro, MWM exhibited significant inhibition of MRSA USA300 and PA27312 growth when compared to negative controls (p ≤ 0.05). Likewise, in-vivo, the MWM-treated wounds exhibited a significant (p ≤ 0.05) bacterial reduction compared to all other treatment groups, especially on days 8 and 12 for both pathogens. MWM demonstrated promise in addressing colonized wounds with biofilms. Additional studies on MWM's benefits and comparisons with existing treatments are warranted to optimize wound care strategies (Adv Wound Care. 2021;10:281).

Integration of multi-omics reveals the important role of the BBS10 gene in reproduction.

Blood samples are easily obtained from sheep. Therefore, blood analysis can be a convenient method for evaluating reproductive traits in sheep by detecting genetic and metabolic changes in the ovary. By combining 167 RNA sequencing (RNA-seq) data and 60 untargeted metabolomics data, this study analyzed the relationship between genes and metabolites in the ovary and blood. The conjoint KEGG enrichment analysis enriched glutathione metabolic pathways both in the ovary and blood. This finding provides an explanation for possible glutathione metabolic processes in the ovary with metabolite exchange in the blood. The metabolite-gene-disease interaction network revealed a correlation between the expression of certain Bardet-Biedl syndrome (BBS) family genes in the ovary and blood. This indicates that BBS family genes, such as BBS10 in sheep blood, could be a potential biomarker for BBS. We investigated the relationship between BBS10 gene expression in the ovary and lambing numbers using whole-genome sequencing data from 450 ewes. Our findings suggest that g.112314188C > G may lead to decreased litter size in ewes carrying the FecB gene. These SNPs could be potential molecular markers for breeding sheep.

Characterization Studies on the sugC Gene of Streptococcus suis Serotype 2 in Adhesion, Invasion, and Virulence in Mice.

The sugC gene of Streptococcus suis (S. suis) is a coding gene for the ATP-binding transporter-associated protein with strong pathogenicity. In order to reveal the effect of the sugC gene on the virulence of S. suis serotype 2, a wild-type strain of TJS75, isolated from fattening pigs' brain tissue samples, was used as a parent strain, and a knockout sugC gene (ΔsugC) and complementary strain (CΔsugC) were successfully constructed via homologous recombination technology. The biological characteristics of TJS75, ΔsugC and CΔsugC were compared and analyzed through growth curves, biochemical characteristics, hemolysis characteristics, cell infection tests and pathogenicity tests on BALB/c mice. The results of the growth characteristic experiments in vitro showed that the plateau stage growth period of ΔsugC was delayed compared to the TJS75 strain, but there was no difference in the total number of bacteria. The biochemical characteristics and hemolysis ability of ΔsugC in sheep blood had no difference compared with TJS75, but its adhesion and invasion abilities in PK-15 cells were decreased. Knockout of the sugC gene had no impact on the expression levels of adhesion-related genes in TJS75 in real-time PCR analysis. In addition, the LD50 of ΔsugC in BALB/c mice was 1.47 × 108 CFU, seven times higher than that of TJS75 (LD50 = 2.15 × 107 CFU). These results illustrate that the deletion of sugC reduced the virulence of TJS75 to BALB/c mice, but its role in the adhesion and invasion of PK-15 cells in this strain needs to be further explored. In summary, this study provides evidence that the sugC gene is a virulence-related gene in the S. suis serotype 2 strain and plays a crucial role in the adhesion and invasion of S. suis. This study lays a foundation for the further exploration of the potential virulence factors and pathogenesis of S. suis.

Validation of the diagnostic accuracy of the 3M™ Petrifilm™ coliform and aerobic count plates to measure colostrum bacterial contamination on Scottish dairy farms

A calf should receive 10–12 % of its bodyweight in high quality colostrum ( >50 g/L immunoglobulin) shortly after birth in order to confer passive immunity, with calves ideally receiving their first feed of colostrum in the first 1–2 hours of life (Godden et al., 2019). It is recommended (not validated) that total counts for bacteria and coliforms in colostrum should not exceed 100,000 colony forming units per mL (CFU/mL) and 10,000 CFU/mL, respectively. This study was conducted between March and September 2023. Eighty-five colostrum samples from 5 commerical dairy farms in Dumfries and Galloway in Scotland were purposively selected. Samples were collected from various stages during the colostrum harvest, storage and feeding process to provide multiple samples with variable (low, medium and high) bacterial contamination. The objective of this study was to validate the diagnostic accuracy of the 3 M Petrifilm™ Coliform Count Plate (CCP) and Aerobic Count Plate (ACP) for colostrum bacteriology. The Petrifilm™ Aerobic Count Plates were compared to 5 % sheep blood agar (SBA) (total bacteria counts, TBC), and the Petrifilm™ Coliform Count Plates were compared to MacConkey plates (MAC) (total coliform counts, TCC) and test sensitivity and specificity were calculated.Colostrum bacterial contamination was highly variable between farms (TBC median and interquartile range= 50,000CFU/mL and 546000CFU/mL; TCC median and interquartile range=100,750CFU/mL and 188,500CFU/mL). Overall correlation between the Petrifilms™ and reference plate measures was moderate (Spearman rho=0.38–0.47). The statistical accuracy of the Petrifilm™ systems and the test sensitivity were improved by lowering the bacterial cutpoint from 100,000CFU/mL to 91,000 CFU/mL for the aerobic counts plates and from 10,000CFU/mL to 1500 CFU/mL for the coliform count plates. The Petrifilm™ system could be used as a monitoring tool to provide quick and statistically accurate results for TBC and TCC but should not be seen as a comprehensive solution for poor colostrum hygiene.

The Effects of Maternal Nutrient Restriction during Mid to Late Gestation with Realimentation on Fetal Metabolic Profiles in the Liver, Skeletal Muscle, and Blood in Sheep.

Poor maternal nutrition during gestation negatively affects offspring growth and metabolism. To evaluate the impact of maternal nutrient restriction and realimentation on metabolism in the fetal liver, skeletal muscle, and circulation, on day 50 of gestation, ewes (n = 48) pregnant with singletons were fed 100% (CON) or 60% (RES) of requirements until day 90 of gestation, when a subset of ewes (n = 7/treatment) were euthanized, and fetal samples were collected. The remaining ewes were maintained on a current diet (CON-CON, n = 6; RES-RES, n = 7) or switched to an alternative diet (CON-RES, RES-CON; n = 7/treatment). On day 130 of gestation, the remaining ewes were euthanized, and fetal samples were collected. Fetal liver, longissimus dorsi (LD), and blood metabolites were analyzed using LC-MS/MS, and pathway enrichment analysis was conducted using MetaboAnalyst. Then, 600, 518, and 524 metabolites were identified in the liver, LD, and blood, respectively, including 345 metabolites that were present in all three. Nutrient restriction was associated with changes in amino acid, carbohydrate, lipid, and transulfuration/methionine metabolic pathways, some of which were alleviated by realimentation. Fetal age also affected metabolite abundance. The differential abundance of metabolites involved in amino acid, methionine, betaine, and bile acid metabolism could impact fetal epigenetic regulation, protein synthesis, lipid metabolism, and signaling associated with glucose and lipid metabolism.

Longitudinal seroepidemiological survey and risk factors for bluetongue virus infection in sheep in the state of Parana, Brazil, from 2014 to 2017.

The Bluetongue disease is an infectious and non-contagious viral disease mainly transmitted through hematophagous vector of the Culicoides genus, to domestic and wild ruminants. The aim of this study was to evaluate the antibodies occurrence, persistence and potential risk factors associated with bluetongue virus infection in sheep flocks in the state of Parana, Brazil. The competitive ELISA test was applied to evaluate 690 blood serum samples from 22 farms in eight mesoregions of Parana in 2014, and 270 sheep blood serum samples from 10 of the 22 previously studied farms in 2017. A questionnaire was applied to evaluate the risk factors associated with BTV infection. In 2014 and 2017, the numbers of seroreactive sheep were found to be 28.26% (195/690) and 41.11% (111/270), respectively, representing 95.45% (21/22) and 90% (9/10) of the flocks. The significant variables considered as risk factors were Culicoides presence (P < 0.0001; OR = 8.83 and 95% CI 4.28-18.22); genealogical record (P < 0.0001; OR = 0.23 and 95% CI 0.12-0.45) and use of sheepfold (P = 0.0208; OR = 0.36 and 95% CI 0.15-0 0.86). It was determined that BTV infection is endemic in Parana and persists in the mesoregions where the climate is favorable to vector proliferation.

Longitudinal detection of prion infection in preclinical sheep blood samples compared using 3 assays

AbstractVariant Creutzfeldt-Jakob disease (vCJD) is a devastating disease caused by transmission of bovine spongiform encephalopathy to humans. Although vCJD cases are now rare, evidence from appendix surveys suggests that a small proportion of the United Kingdom population may be infected without showing signs of disease. These “silent” carriers could present a risk of iatrogenic vCJD transmission through medical procedures or blood/organ donation, and currently there are no validated tests to identify infected asymptomatic individuals using easily accessible samples. To address this issue, we evaluated the performance of 3 blood-based assays in a blinded study, using longitudinal sample series from a well-established large animal model of vCJD. The assays rely on amplification of misfolded prion protein (PrPSc; a marker of prion infection) and include real-time quaking-induced conversion (RT-QuIC), and 2 versions of protein misfolding cyclic amplification (PMCA). Although diagnostic sensitivity was higher for both PMCA assays (100%) than RT-QuIC (61%), all 3 assays detected prion infection in blood samples collected 26 months before the onset of clinical signs and gave no false-positive results. Parallel estimation of blood prion infectivity titers in a sensitive transgenic mouse line showed positive correlation of infectivity with PrPSc detection by the assays, suggesting that they are suitable for detection of asymptomatic vCJD infection in the human population. This study represents, to our knowledge, the largest comparison to date of preclinical prion detection in blood samples from a relevant animal model. The outcomes will guide efforts to improve early detection of prion disease and reduce infection risks in humans.

Dolosigranulum savutiense sp. nov., isolated from human upper respiratory samples collected in Botswana.

Four strains (MSK211, MSK294T, MSK312, MSK433) of a novel Dolosigranulum species were cultured from nasopharyngeal swabs collected from mother-infant dyads in southern Botswana. These strains grew optimally on tryptic soy agar with 5% sheep blood solid medium and in fastidious bacteria broth. Colonies on tryptic soy agar with 5% sheep blood agar appeared grey or white with a flat, smooth surface and variable alpha haemolysis. Cells were Gram-positive, non-spore-forming, non-motile cocci that lacked catalase or oxidase activity. Major fatty acids were C16 : 0 (palmitic acid), C18 : 1 ω9c (oleic acid), and C18 : 0 (stearic acid). Analyses of 16S rRNA gene sequences identified these strains as belonging to the genus Dolosigranulum (family Carnobacteriaceae), which currently contains only a single validly published species (Dolosigranulum pigrum). Whole-genome sequencing revealed that the genomes of these strains are 1.98-2.07 Mbp in size and have a G+C content of 39.6-39.9 mol%. Comparisons of these genomes to publicly available genomes of D. pigrum yielded average nucleotide identities and in silico DNA-DNA hybridization values of 92.3-92.9% and 49.1-51.4%, respectively. These results indicate that these strains represent a novel species of Dolosigranulum, for which we propose the name Dolosigranulum savutiense sp. nov., with the type strain MSK294T (=DSM 117171T=JCM 36673T).

Prevalence and antimicrobial susceptibility of Streptococcus equi isolated from horses in Santa Catarina state, Southern Brazil.

The objective of this study was to determine the prevalence of Streptococcus equi (S. equi subsp equi and S. equi subsp zooepidemicus) in the state of Santa Catarina and evaluate the antimicrobial susceptibility of the isolates. For this, 420 nasal swab samples were collected from randomly selected horses. Isolation and phenotypic characterization of the bacteria were performed by sowing on 5% sheep blood agar, followed by analysis of morphotinctorial characteristics and biochemical analysis. To differentiate the main beta-hemolytic Streptococcus in horses, the fermentation profiles of the sugar's lactose, maltose, sorbitol, and trehalose were used, which were confirmed at the subspecies level by the PCR technique. The antimicrobial susceptibility panel was defined by the disk diffusion method, testing 13 antimicrobials from ten different classes, all regularly used in equine medical clinics, followed by the calculation of the multiple antimicrobial resistance index. Ten strains of S. equi were isolated, with a prevalence of 2.38% (10/420). Of the total positive samples, 3% (3/10) were confirmed as belonging to S. equi subsp equi and 70% (7/10) were confirmed as belonging to S. zooepidemicus. Multidrug resistance was observed in 60% (6/10) of isolates. The antimicrobial with the greatest resistance was clindamycin with 70% (7/10), followed by beta-lactams, with 40% (4/10) resistance to penicillin and 30% (3/10) to ceftiofur. The isolates were 100% (10/10) sensitive to gentamicin, chloramphenicol, levofloxacin, and vancomycin. This was the first study carried out in the state, and based on these data, it can be said that Santa Catarina has a low prevalence of S. equi and the presence of multi-resistant strains of S. equi was confirmed in the equine herd in Santa Catarina.

Influence of drying time on the removal of blood from medical devices

BackgroundWhen processing surgical instruments after use, the safe, residue-free removal of blood and blood-containing soiling is one of the most important tasks. There are recommendations from various working groups regarding the ideal time frame for cleaning used instruments in order to ensure safe disinfection and sterilisation and avoid adverse effects. These are generally based primarily on practical experience and there is little systematic work on this topic. AimIn the present study, cleaning experiments with test specimens previously contaminated with sheep's blood were carried out and in this way the effects of the drying time of whole blood on the results of the subsequent cleaning were examined. MethodsReflecting practice, both visual and spectroscopic methods were used to quantify residual protein. The experimental results were evaluated both as a function of the drying time and the residual moisture of the blood. FindingsIt was found that drying blood is particularly difficult to remove within the first 1-2 hours. In this phase, in which the blood is coagulated but not yet completely dried, considerably more protein residues remained on the test specimens after cleaning than after longer standing times. ConclusionsIt was concluded that there is a time frame for the removal of blood residues in which optimum cleaning results can be expected. As a consequence, there are also standing times that are disadvantageous for reprocessing. Based on the experimental data, it was deduced that this optimum time is either directly after contamination or in the range of > 3 h and < 24 h after soiling.

Induction and comparative resuscitation of viable but nonculturable state on Vibrio parahaemolyticus serotypes O3:K6 and O1:K25.

Vibrio parahaemolyticus, an important food-borne pathogens found to be associated with seafoods and marine environs. It has been a topic of debate for many decades that most pathogens are known to enter a viable but nonculturable (VBNC) state under cold temperature and nutrient limited conditions. The present study examined the time required for the induction of VBNC state and the revival strategies of both the endemic O3:K6 and O1:K25 sporadic strains of V. parahaemolyticus. The results revealed that V. parahaemolyticus survived even after 55 days of incubation in nutrient starved media such as phosphate buffered saline (PBS) and Coastal Water (CW) and could be recovered by temperature upshift method, and compared the resuscitation using Dulbecco's Modified Eagle Medium (DMEM), sheep blood serum, chitin flakes with live Artemia salina, and the results suggests that chitin plays a significant role in regulating the VBNC state. It was also confirmed by Confocal Laser Scanning Microscopy (CLSM) and Scanning Electron Microscope (SEM) analysis that VBNC cells can alter their morphology to coccoid forms in order to survive in most extreme nutrient limited environment. Further data on the promoting factors and the exact mechanism that resuscitate VBNC V. parahaemolyticus in cold natural environments and frozen foods are needed to perform a robust risk assessment.

Microbial Contamination in Contact Lenses, Lens Care Solutions, and Accessories Among Asymptomatic Soft Contact Lens Users.

Objective This study aimed to evaluate microbial contamination of contact lenses (CL) and their accessories among asymptomatic lens users and identify behavioral risk factors that might exacerbate the said contamination. Methodology Ninety-five asymptomatic soft CL users were recruited. In total, 380 samples were collected from the inner surface of lenses, the base of lens cases, the tip of the multipurpose solution bottle, and the solution itself. All swabs with samples were inoculated onto Columbia 5% sheep blood agar, MacConkey agar, Pseudomonas agar with cetrimide, and Sabouraud dextrose agar. Blood agar, MacConkey agar, and Pseudomonas agar with cetrimide were incubated at 37 °C for 24-48 hours. Fungal growth was investigated on Sabouraud dextrose agar, incubated at 25 °C, and examined daily for three weeks. Microscopic examination, culture-based methods, and biochemical tests were used to identify isolated microorganisms. A self-administered questionnaire on compliance with care and hygiene procedures was completed by each participant. Results The overall microbial contamination of tested samples was 38.7%. The most frequently contaminated items were lens cases (59, 62.1%), followed by bottles (44, 46.3%) and lenses (35, 36.8%). Meanwhile, the lowest incidence of contamination was seen in lens multipurpose solutions (9, 9.5%). The predominant microorganisms recovered were Coagulase-negative Staphylococci (CoNS) (94, 64%) and Gram-positive rods (29, 19.7%). Other identified potential pathogens were Staphylococcus aureus(11, 7.5%), Pseudomonas aeruginosa(5, 3.4%), Escherichia coli(1, 0.7%), andCandida albicans(2, 1.4%). The questionnaire revealed that contact lens users aged 18 to 20 showed a lack of compliance with proper hygienic care for contact lens maintenance. Risk factors such as male gender, smoking, showering, or swimming while wearing CL were related to microbiological contamination in at least one of the samples (P > 0.05). Conclusions The highest degree of contamination with highly virulent pathogens was determined in lens cases owing to insufficient lens care practices among study participants. Noncompliance with the lens cleaning procedures can lead to microbial colonization of the lens and its accessories, prompting inflammatory events in the eyes in the future.

Genetic analysis of platelet-derived growth factor receptor-like gene (PDGFRL) polymorphism and melanin traits in Lanping black-boned sheep (Ovis aries)

Abstract. In the intricate tapestry of Yunnan Province's biodiversity, the Lanping black-boned sheep (LPBB) emerges as a captivating enigma, distinguished by its profound melanin pigmentation adorning both its skin and its internal organs. Initially cataloged in the 1950s within the confines of Lanping County, this exceptional mammalian species presents a scarcity and uniqueness that extends beyond its geographic origins. Here, we collected 100 blood samples from Lanping black-boned sheep along with 50 samples each from Lanping normal sheep (LPN) and Huize normal sheep (HZN), all sourced from Yunnan Province. Our investigation focused on the association between the platelet-derived growth factor receptor-like gene (PDGFRL) polymorphism and the distinctive melanin characteristics observed in Lanping black-boned sheep. Utilizing UV–visible spectrophotometry, we assessed the melanin indexes present, such as tyrosinase activity and true melanin in the sheep blood, and the results demonstrated a significant elevation in melanin indexes for Lanping black-boned sheep compared to the control group (P&lt;0.05). We also identified three synonymous mutation sites within a partial 1128 bp exon fragment of the gene-encoding PDGFRL (EX2-G408A, EX5-T184C, and EX5-G222T). Notably, Lanping black-boned sheep, harboring genotypes GG, TT, and GG at these specific sites, showcased a pronounced surge in tyrosinase activity, eumelanin / total melanin ratios, and plasma colorimetric values when contrasted with the control group (P&lt;0.05). The discernment of GG, TT, and GG as the prevailing genotypes at their respective genetic loci in Lanping black-boned sheep heralds a breakthrough in our understanding of the genetic markers associated with black pigmentation. However, all three loci are silent mutations and do not alter the phenotypic changes. Whether they affect changes in melanin content through other metabolic pathways requires further study. In conclusion, the PDGFRL gene was silenced by mutations in our study and affected blood melanin levels. However, the gene did not undergo a missense mutation that altered the phenotypic changes, and the exact channel through which the changes in melanin content were affected needs to be further verified.

Evaluation of gradient diffusion in vitro susceptibility testing of Aerococcus urinae

ObjectiveAerococcus urinae antimicrobial susceptibility testing can be performed via broth microdilution with Mueller-Hinton broth supplemented with lysed horse blood. We sought to compare this with the commonly used gradient diffusion method. MethodsWe compared broth microdilution with Mueller-Hinton broth supplemented with lysed horse blood and gradient diffusion via Mueller-Hinton agar supplemented with sheep blood for 190 A. urinae isolates against 16 antimicrobials. ResultsNo antimicrobials demonstrated more than 90% essential and categorical agreement, and fewer than 3% demonstrated major and very major error rates. Trimethoprim-sulfamethoxazole demonstrated an 81% major error rate and ceftriaxone demonstrated a 76% very major error rate. Agar dilution with lysed horse blood was performed for trimethoprim-sulfamethoxazole against 94 isolates and showed 100% susceptibility, consistent with previous studies. ConclusionsGiven its limitations in detecting resistant strains, our findings cannot support the routine use of gradient diffusion with Mueller-Hinton agar supplemented with sheep blood for A. urinae in lieu of the Clinical and Laboratory Standards Institute method. Our results suggest that A. urinae is usually susceptible to penicillin, linezolid, tetracycline, and vancomycin. Future studies should evaluate alternative testing methods for clinical microbiology laboratories.

Evaluating the Bias of Two Point-of-Care Glucometers for Calves and Ewes: Awareness for Ruminant Practitioners

(1) Background: Multiple point-of-care (POC) glucometers are in use in veterinary medicine, but few are compared to each other. This leaves the potential for clinicians to be unaware of the effect of bias when comparing results from different POC glucometers. (2) Methods: Samples from healthy calves and ewes were simultaneously compared with two POC veterinary glucometers, the Precision Xtra and the AlphaTrak2, under both the “canine” and “feline” settings. The results of each sample were statistically analyzed with linear regression and Bland–Altman analysis. (3) Results: 170 samples from healthy calves and 108 samples from healthy ewes were available for comparison. Calves: The AT2 consistently overestimated blood glucose concentrations when compared to the PX device with the calves. Correlationt with the PX was r = 0.8496 (canine setting) and r = 0.8861 (feline setting). Both the canine and feline settings demonstrated a consistent bias (41.11 and 33.64 mg/dL, respectively). Ewes: The AT2 consistently overestimated blood glucose concentrations when compared to the PX device with the ewes. Correlation with the PX was R = 0.4710 (canine setting) and R = 0.7269 (feline setting). Both the canine and feline settings demonstrated a consistent bias (21.23 and 14.54 mg/dL, respectively). (4) Clinicians should be aware of the potential for consistent bias when evaluating calf and sheep blood glucose concentrations as the AT2 device, at both settings, overestimated blood glucose compared to the previously validated PX. This reliability appears to change when the values are farther from the normal ranges, which should be considered when making clinical decisions based on data from these devices.

Profile characteristics of sensitivity to antimicrobial chemotherapeutic drugs of Corynebacterium jeikeium isolated in explosive mine injuries of the limbs

Background. From the moment of gunshot wounds, the microbiological landscape depends on the provision of medical care at each evacuation link. Effective antibacterial therapy relies on choosing the right antibiotics based on the pathogen’s sensitivity to them. This approach aims to prevent infectious complications of combat trauma, and it requires continuous microbiological monitoring of circulating microbial strains. Purpose – to study the profile of microorganisms isolated from mine-explosive injuries of the limbs of patients who were admitted to the last stage of medical care in the regional centre of the city of Lviv. To determine the antibiotic susceptibility of the isolated nosocomial pathogen C. jeikeium to antimicrobial chemotherapeutic agents. Materials and Methods. The strains of microorganisms isolated from gunshot wounds of the limbs of 20 male casualties, aged 20 to 50 years upon admission to the Lviv Regional Medical Facility in 2024, were investigated. The collection of material was carried out using sterile swabs in a transport nutrient medium. The material was inoculated onto blood agar, Endo medium, CHROMID® P. aeruginosa Agar, MPA, and incubated at 37°C for 24 hours. Biochemical identification was carried out using the Enterotest 24 and Nefermtest (Lachema, Czech Republic). The cultivation of C.jeikeium was carried out on sheep blood agar, with the purpose of accelerating growth, Tween 80 was used. It was identified based on morpho-tinctorial and cultural properties, and its biochemical activity was studied through the fermentation of glucose and galactose. Considering the fact that the predominant contaminants of combat wounds were 8 strains of C.jeikeium, the sensitivity of these isolates to antimicrobial chemotherapeutic agents was determined using the disc diffusion method with standard discs EUCAST. The sensitivity of the above-mentioned isolates was determined using the disc diffusion method with the use of standard discs. At evaluation of resistance profiles was guided by the EUCAST standard. Results. As a result of the operation, 24 pure cultures were isolated and identified to the species level. The sensitivity of selected isolates of C.jeikeium to vancomycin and linezolid was set at 100%. Sensitivity to tetracycline was 25%, gentamicin and clindamycin – 12.5% each. Given that the sensitivity of 8 isolates of C.jeikeium turned out to be similar; that the majority of cases of infection with this microorganism have a pronounced hospital character, we assume that we have isolated a nosocomial pathogen. Conclusions. Summarizing the results of the study and comparing with the data of other authors, regarding the monitoring of the microbiological landscape of wounds of the limbs, we state that in our study the frequency of detection of P.aeruginosa was – 8.3%, K.pneumoniae – 20.8%, not a single case of isolation of A.baumanii. The most frequently isolated microorganisms were: C.jeikeium – 8 strains (33.3%) with similar sensitivity to antimicrobial chemotherapeutic drugs, which may indicate the isolation of a nosocomial pathogen. The drugs of choice for C.jeikeium elimination are vancomycin and linezolid. After obtaining the result of sensitivity to antibiotics, it is possible to prescribe tetracy cline, gentamicin, clindamycin.

Molecular detection and characterization of Anaplasma ovis, Theileria ovis, and Theileria lestoquardi in sheep and goats in Luxor, Egypt

BackgroundTick-borne diseases cause economically significant losses to animal production globally, and anaplasmosis and theileriosis are associated with the greatest losses. However, the spread of the relevant pathogens in flocks of domesticated animals in southern Egypt is little understood. Accordingly, in this study, we aimed to determine the prevalences of Anaplasma ovis, Theileria ovis, and Theileria lestoquardi in southern Egyptian sheep and goats through blood tests, and to make a molecular characterization of the A. ovis detected in sheep targeting a specific gene.ResultsWe collected blood samples collected from 300 sheep and goats (n=150 /species) in Luxor Province in southern Egypt, and analyzed them for the presence of A. ovis, T. ovis and T. lestoquardi with screening by conventional and nested PCR targeting the msp4 and msp5, 18S rRNA, and merozoite surface protein genes. For A. ovis 140/300 samples (46.66%) were positive overall, with 90/150 (60%) and 50/150 (33.33%) positive samples in sheep and goats, respectively. Two major surface protein genes of A. ovis, msp4 and msp5, were sequenced using DNA extracted from sheep and goat blood samples, for phylogenetic analysis and genotyping. The msp4 gene sequence revealed no significant genetic diversity, to contrast to data on A. ovis strains from other countries. For T. lestoquardi, 8/150 (5.33%) samples were positive in sheep, but no samples were positive in goats (0%). For T. ovis, 32/150 (21.33%) samples were positive in sheep, but no samples were positive in goats (0%). Sequencing targeting the merozoite surface protein gene for T. lestoquardi and the small subunit ribosomal RNA gene for T. ovis revealed no significant genetic diversity in the study, another contrast to data on A. ovis strains from other countries.ConclusionThis study provides valuable data on phylogenetic and molecular classifications of A. ovis, T. ovis and T. lestoquardi found in southern Egyptian sheep and goats. It also represents the first report on detection and molecular characterization of T. lestoquardi in southern Egyptian sheep based on the specific merozoite surface protein gene, thus providing valuable data for molecular characterization of this pathogen in southern Egypt.