Phenotypic and genotypic characterization of erythromycin-resistant Staphylococcus aureus isolated from bovine subclinical mastitis in Egypt.

Background and Aim: Subclinical mastitis (SCM) caused by erythromycin-resistant Staphylococcus aureus is a significant disease in lactating animals. Therefore, it is crucial to understand the genetic factors contributing to erythromycin resistance in S. aureus. This study aimed to estimate the prevalence of S. aureus in milk from subclinical mastitic cattle and buffaloes and tank milk samples as identified by probe-based real-time polymerase chain reaction (PCR) and the genotypic assessment of macrolide and erythromycin resistance profiles, as well as to analyze the phylogenetic relatedness of our local isolates of S. aureus. Materials and Methods: In total, 285 milk samples were analyzed using the California mastitis test to detect SCM. Milk samples were cultured on different specific Staphylococcus media. The presence of S. aureus was confirmed by Gram staining, the catalase and coagulase tests, the detection of hemolytic activity, DNase agar testing, and biofilm activity in Congo red medium. The genotypic identification of S. aureus (nuc) was performed. The determinants of erythromycin (ermA, ermB, ermC, and ermT) and macrolide resistance (msrA) were screened in all isolates. DNA sequencing of our local isolates of S. aureus was used to analyze their phylogenetic relatedness. Moreover, histopathological examination of tissue specimens of mammary gland was performed. Results: The S. aureus positivity rates were 36.4%, 48.8%, and 63.6% in cattle, buffalo, and bulk tank milk, respectively. Probe-based real-time PCR molecularly confirmed all 62 S. aureus isolates. Thirty-one isolates were subjected to PCR to create profiles of their genotypic erythromycin resistance. ermA, ermB, ermC, and ermT were present in 5 (8%), 26 (41.9%), 18 (29%), and 15 (24.1%) S. aureus isolates, respectively. Moreover, msrA was found in three (4.8%) strains. Eight PCR products were produced using standard PCR for DNA sequencing. Multiple sequence alignment, phylogenetic tree construction, and analysis of nuc in S. aureus revealed a high degree of homology (100%) with S. aureus strains isolated from milk in cases of bovine mastitis in India and Kenya. Histological analysis of udder tissues revealed extensive aggregation of mononuclear inflammatory cells in the interstitial connective tissue, primarily lymphocytes, and macrophages. Conclusion: This study showed a high prevalence of erythromycin resistance in S. aureus isolates. This information is vital for controlling mastitis and the spread of resistance genes between bacterial strains and hosts. Moreover, the probe-based real-time PCR approach is helpful for the rapid screening of S. aureus isolates and the consequent efficient treatment and control of S. aureus mastitis.

Molecular detection and identification of Babesia bovis and Trypanosoma spp. in one-humped camel (Camelus dromedarius) breeds in Egypt.

BACKGROUND AND AIM: Camels are a unique source of milk and meat, which helps recover from several diseases that affect humans worldwide. In Egypt, one of the great obstacles for this industry is tick-borne diseases. This study aimed to characterize blood parasite infections, such as Babesia (B.) bovis and Trypanosoma (T.) spp. in one-humped camel (Camelus dromedarius) (n=142) breeds in Halayeb and Shalateen, Egypt, through phylogenetic analysis. MATERIALS AND METHODS: The prevalence of B. bovis and Trypanosoma spp. was identified in camels using polymerase chain reaction (PCR) assays targeting the Rhoptry-Associated Protein-1 and internal transcribed spacer 1 genes, respectively. A nested PCR technique was conducted to detect B. bovis. At the same time, KIN multispecies PCR assay was employed to diagnose and classify trypanosome DNA in camels. RESULTS: B. bovis was detected in 4/142 camels with an infection rate of 2.81%. Sequencing and phylogenetic analyses revealed that the strain of B. bovis isolated from this population was closely related to strains isolated from Argentine, the United States, and Brazil. Moreover, Trypanosoma evansi was detected in 8/142 camels with an infection rate of 5.63%. Sequencing and phylogenetic analyses revealed that this isolated strain T. evansi was closely related to Trypanosoma theileri detected from cattle in Brazil. CONCLUSION: The obtained data indicated the existence of B. bovis and T. evansi in camels from two provinces of Egypt. The obtained findings have economic significance and reflect the importance of implementing effective prevention and control methods across Egypt to reduce the incidence of B. bovis and T. evansi in camels.

Congestive heart failure in cattle; etiology, clinical, and ultrasonographic findings in 67 cases.

BACKGROUND AND AIM: Congestive heart failure (CHF) is a clinical disorder that results from cardiac dysfunction with subsequent fatal outcomes in most cases. Several diseases are incriminated in occurrence of CHF. Therefore, the aims of this study were to identify CHF etiology and associated clinical findings in 67 cows and to investigate the relationship between CHF and the other body organs using ultrasonographic examination. MATERIALS AND METHODS: Sixty-seven cows affected by CHF admitted to the clinic with a history of loss of appetite, decrease in milk production, constipation, and brisket edema were thoroughly investigated clinically and ultrasonographically. In addition, ten apparently healthy cows were used as a control group. RESULTS: Clinically, cows with CHF manifested jugular engorgement and pulsation (88.1%), brisket and/or intermandibular edema (77.6%), and muffled heart sounds (76.1%). Based on the ultrasonographic examination, traumatic pericarditis (82.1%) was the most prevalent etiology of CHF. Extracardiac etiology of CHF identified were exudative pleurisy (10.4%) and mediastinal abscesses (7.5%). Hepatomegaly (88.1%) and pleural effusion (61.2%) were the most documented consequences. CONCLUSION: Both cardiac and extracardiac diseases could be associated with CHF in cattle. Ultrasonographic changes in liver and pleura secondary to CHF were the most common findings. Ultrasonography is a good tool for the diagnosis of cardiac and extracardiac etiologies of CHF in cattle.

Diagnostic performance of ultrasonography in clinical management of dairy cattle identified with left-sided ping sounds.

OBJECTIVES: Left displaced abomasum (LDA) is a common postparturient condition of high yielding dairy cattle. The diagnosis of LDA is challenging and has historically been based on findings that are not specific to the condition. The objective of the current study was to investigate the diagnostic performance of ultrasonography (USG) in the clinical management of dairy cows identified with left-sided ping sound postpartum. MATERIALS AND METHODS: Cows with reduced appetite postpartum and had audible left-sided ping sounds on abdominal auscultation were eligible to be prospectively recruited onto the study. The results of clinical findings and abdominal USG were recorded along with milk ß-hydroxybutyrate levels, pH levels of abomaso/rumenocentesis samples, and findings on exploratory laparotomy. The diagnostic performance of USG and other clinical investigations was assessed by calculating the test sensitivity and specificity using exploratory laparotomy as a gold standard test. RESULTS: A definitive diagnosis of LDA was made in 23 cows, 8 cows were diagnosed with peritonitis, and 4 cows with frothy tympany. The USG findings that were consistent with LDA were present in all cattle diagnosed with LDA at exploratory laparotomy. The USG findings over the past three intercostal space characteristics of LDA, however, were also present in five cases subsequently diagnosed with peritonitis and in all cases diagnosed with frothy tympany on exploratory laparotomy. The pH of abdomaso/rumenocentesis samples yielded the highest diagnostic accuracy (97.14%) as a single test in the current study. CONCLUSIONS: USG over the left abdominal wall despite being a highly sensitive test for the diagnosis of LDA has limitations as a diagnostic tool due to suboptimal specificity.

Aflatoxicosis in cattle: clinical findings and biochemical alterations.

Aflatoxicosis is a serious health condition resulted from aflatoxin (AF)-producing fungi. Major health threats resulted from AFs and reflect on the livestock industry with great economic losses. There are limited scientific evidences concerning the AFs in ruminant, therefore it is important to evaluate AFs health hazards in cattle. Here, we investigate biochemical, oxidative stress, and postmortem changes associated with unexpected acute bovine aflatoxicosis. Seventy-two cattle were suffered from aflatoxisocis. Depression and inappetence were predominant clinical findings of the diseased animals. Analysis of feedstuffs revealed presence of aflatoxin B1 (AFB1). The AF-intoxicated animals showed a significant increase in alanine amino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP), serum creatinine (SCr), catalase (CAT), and malondialdehyde (MDA) levels. Moreover, a significant decrease in total protein (TP), magnesium (Mg), and reduced glutathione (GSH) were also seen. Hepatomegaly, enlarged gallbladder as well as congestion of the intestine and kidney were observed. This study elucidates the critical and constructive measurements needed for the prevention of the AFs hazardous effects to livestock for the future control of AF outbreaks. Conducting series of diagnostic assays reflect the marked health condition alterations in the biochemical and antioxidant status of the AF-intoxicated cattle.

Isolation and characterization of vaginal Lactobacillus spp. in dromedary camels (Camelus dromedarius): in vitro evaluation of probiotic potential of selected isolates.

Lactobacillus spp. is one of the beneficial lactic acid producing microbiota in the vagina, which is important for a healthy vaginal environment. However, little is known about vaginal Lactobacillus in dromedary camels (Camelus dromedarius). Therefore, this study aimed to isolate vaginal lactic acid bacteria (LAB) in dromedary camels and to study the probiotic potential of selected isolates. A total of 75 vaginal swabs were collected from pluriparous, non-pregnant, non-lactating dromedary camels. The LAB were isolated using deMan, Rogosa and Sharpe broth and agar media. Suspected LAB isolates were subjected to catalase testing and Gram staining and examined for indole production, nitrate reduction, hemolytic activity, cell surface hydrophobicity, auto- and coaggregation, antibacterial activity and characterized by 16S rRNA amplification and sequencing. Eighteen LABs were isolated from the 75 vaginal swabs. Among the 18 LAB isolates, six were Lactobacillus plantarum, eight were Lactobacillus fermentum, and four were Lactobacillus rhamnosus. None of the LAB isolates was hemolytic and only four LAB were H2O2 producing. The percentage of hydrophobicity ranged from 0% to 49.6%, 0% to 44.3% and 0% to 41.6% for hexadecane, xylene and toluene, respectively. All isolates showed higher (P < 0.05) autoaggregation after 24 h of incubation compared to 4 h. Furthermore, all LAB showed higher coaggregation (P < 0.05) and antimicrobial activity toward Staphylococcus aureus than to Escherichia coli. All LAB isolates were vancomycin resistant and sensitive to streptomycin, erythromycin, kanamycin and chloramphenicol. Only, three LAB isolates were resistant to tetracycline. The dromedary camel vaginal LAB isolates exhibited varying degrees of in vitro probiotic properties tested in this study and showed promising activity against the most common bacterial causes of endometritis in dromedary camels. Further investigation of the in vivo effect of these isolates is warranted.

Use of procalcitonin, neopterin, haptoglobin, serum amyloid A and proinflammatory cytokines in diagnosis and prognosis of bovine respiratory disease in feedlot calves under field conditions.

Bovine respiratory diseases (BRD) have long been considered a serious problem that causes major economic losses in feedlot calves (FC). This study aimed to determine the diagnostic and prognostic effect of selected biological markers including, procalcitonin (PCT), neopterin (NP), proinflammatory cytokines (IL-1ß, IL-8, TNF-α, IF-γ), haptoglobin (HP) and serum amyloid A (SAA) on FC with BRD under field conditions. Sixty-nine FC that were identified to be infected with Mannheimia haemolytica and Histophilus somni and had different clinical respiratory signs (diseased group) were selected for this study. In addition, 20 healthy FC have been selected as a control group. We have detected higher serum levels of PCT, NP, HP, SAA, IL-1ß, IL-8, TNF-α and IF-γ in diseased FC group compared with the control group. All tested markers revealed a high level of discrimination between BRD infected FC and healthy ones (AUC > 0.90). Moreover, the obtained data showed a high degree of prognostic accuracy for PCT, NP, IL-8, HP, IF-γ and IL-1ß in predicting treatment response of FC with BRD at the selected thresholds (AUC = 0.99, 0.99, 0.97, 0.93, 0.88 and 0.82, respectively). Significant inhibition was observed for the selected biochemical markers in treated FC 7 days post-treatment. In conclusion, this study showed that BRD in FC was associated with significant alterations in serum APPs, proinflammatory cytokines, PCT and NPT levels. Furthermore, it demonstrated that these serum biomarkers are much higher in FC with BRD compared to recovered ones. Our data suggest that the measurement of PCT, NPT, APPs and cytokines together with the clinical examination may be a useful diagnostic and prognostic tool for assessment of FC naturally infected with M. haemolytica and H. somni.

The Contribution of Specific and Nonspecific Biomarkers in Diagnosis of Equine Gastric Ulcer Syndrome (EGUS) Under Field Condition.

The aim of this study is to investigate the diagnostic efficiency of gastrin, pepsinogen, proinflammatory cytokines (TNF-α, IL-6), and oxidative stress biomarkers in horses with equine gastric ulcer syndrome (EGUS). Thirty horses diagnosed with gastroscopic EGUS and 15 clinically healthy horses were selected for this study. The serum levels of gastrin, pepsinogen showed nonsignificant changes in horses with EGUS when compared with healthy horses. The serum levels of TNF-α, IL-6 revealed a significant increase in horses with EGUS when compared with healthy ones. Oxidative stress is evident in horses with EGUS in comparison with healthy horses as detected by higher levels of malondialdehyde (MDA) and decreased serum levels of total antioxidant capacity (TAC), Superoxide dismutase (SOD), glutathione (GSH), and nitric oxide (NO). MDA and TNF-α showed better sensitivity and specificity than IL-6 in distinguishing horses with EGUS from control horses. Conclusively, examination of serum gastrin and pepsinogen levels had a limited value in diagnosis of EGUS in horses under investigation. Moreover, this study showed that oxidative stress is evident in horses with EGUS. Higher levels of TNF-α and IL-6 indicate their role in EGUS pathogenesis in horses. Finally, MDA, TNF-α, and IL-6 could be used as biological markers for preliminary screening of horses with EGUS. Gastroscopy still accredited as the "gold standard" for diagnosis EGUS.

Acute phase proteins, proinflammatory cytokines and oxidative stress biomarkers in sheep, goats and she-camels with Coxiella burnetii infection-induced abortion.

Acute phase proteins (APPs) and oxidative stress are helpful markers in diagnosis of several infectious diseases. APPs, proinflammatory cytokines and oxidative stress markers were evaluated for their role in the diagnosis of naturally acquired Coxiella burnetii (Q fever) associated with abortion in sheep, goats and she-camels. Blood, aborted materials and vaginal swabs were collected from mixed herds in the Eastern Province of Saudi Arabia. Antioxidant biomarkers showed significant decline in cases of abortion compared to control animals at delivery time. The correlation between disease status and all parameters ranged from moderate to high. The APPs, cytokines and the oxidative stress marker malondialdehyde (MDA) displayed a high degree of distinction between aborted sheep and goat and normal delivered animals (AUC > 0.90). However, only MDA showed a high degree of differentiation (AUC > 0.90) between aborted she-camels and normal delivered controls. In conclusion, results from our study allow us to recommend using APPs, cytokines and oxidative stress markers as an additional tool for diagnosis of naturally occurring C. burnetii infection in sheep, goats and she-camels. However, it does not replace standard procedures for detection of C. burnetii.

The first molecular detection of Clostridium perfringens from pneumonic cases associated with foot and mouth disease in cattle and buffalo in Egypt.

Panting syndrome and respiratory infection have been recorded in complicated cases of foot and mouth disease (FMD) in cattle. However, investigations on the causative agents of respiratory disease in such cases are scarce. In this study, 30 animals (13 buffalo and 17 cattle) suffering from respiratory distress associated with signs of FMD were examined. Serum samples were collected and FMD infection was confirmed. Bacteriological examination of lungs from eight necropitized cases revealed the presence of C. perfringens. Multiplex polymerase chain reaction (mPCR) was performed on the positive samples followed by sequencing analysis. The alpha toxin gene (plc) of C. perfringens was identified in six cases. The present investigation highlights the role of clostridial infection as a complication of FMD in cattle and buffalo. This is the first report identifying the C. perfringens toxins from lung of animals with respiratory distress associated with FMD infection.