Ziehl-Neelsen Research Articles

Potential zoonotic agents in sea turtles: Chlamydia species and Listeria monocytogenes

Abstract Background Sea turtles are well recognized sentinels of marine environment as well as carrier of pathogens and contaminants. Chlamydial species have been described in reptiles, but scarce data regarding sea turtles are available in literature (Pace et al., 2022). Hitherto, only one case of listeriosis has been described in marine reptiles (Di Renzo et al., 2022). This study reports the detection of Chlamydia species and Listeria monocytogenes in loggerhead sea turtles (Caretta caretta), suggesting the role of these animals as carriers of potential zoonotic agents. Methods In 2022, a total of 11 sea turtles found stranded along Emilia-Romagna coast, Northwestern Adriatic Sea, were subjected to post mortem investigation. Cloacal swabs were collected from 10 of the investigated animals and screened for Chlamydiaceae family by qPCR targeting 23S rRNA gene, followed by confirmation via 16S rRNA fragment sequencing. For positive samples, Chlamydia species identification was performed by species-specific qPCR for C. abortus, C. pneumoniae, C. pecorum, C. psittaci. One subject showed internal organs disseminated with multiple nodular lesions that were sampled for histopathology analyses, alongside microbiological examination. The cultured isolates were identified by MALDI-TOF (MALDI Biotyper; Bruker Daltonics Inc). Results In six out of 10 cloacal swabs Chlamydiaceae DNA was demonstrated by qPCR and confirmed by sequencing of 16S rRNA, but species identification was not achieved. In the turtle with nodular lesions, histopathology revealed the presence of granulomas: the Ziehl-Neelsen stain was negative for acid-fast organisms. L. monocytogenes was isolated from heart and liver and listeriosis was diagnosed. Conclusions The results highlight how sea turtles can be carriers, symptomatic or not, of pathogens relevant for public health. Given the zoonotic potential of these microorganisms, particular precautions are recommended especially for professionals handling these animals. Key messages • Sea turtles can play an active role as carriers of possibly pathogenetic microorganisms. • Further studies of molecular identification of Chlamydia spp. from sea turtles are needed in atypical strains.

Monitoring the immune response of macrophages in tuberculous granuloma through the expression of CD68, iNOS and HLA-DR in naturally infected beef cattle

Bovine tuberculosis still represents a universal threat that creates a wider range of public and animal health impacts. One of the most important steps in the pathogenesis of this disease and granuloma formation is the phagocytosis of tuberculous bacilli by macrophages. Mycobacteria replicate in macrophages, which are crucial to the pathophysiology of mycobacterial infections; however, scarce information is available about the dynamics of the granuloma-stage immunological response. Therefore, immunohistochemistry was used in this work to evaluate the expression of CD68, iNOS, and HLA-DR in different stages of TB granulomas from naturally infected cattle with tuberculosis. Two thousand, one hundred and fifty slaughtered beef cattle were examined during the period from September 2020 to March 2022. Sixty of them showed gross tuberculous pulmonary lesions and samples were collected from all of them for histopathological examination, Ziehl-Neelsen (ZN) staining, and bacteriological culturing. Selected samples that yielded a positive result for ZN and mycobacterial culturing were subjected to an immunohistochemical study of CD68, iNOS, and HLA-DR expression by macrophages according to granuloma stages. Immunohistochemical analysis revealed that the immunolabeling of CD68+, iNOS+, and HLA-DR+ macrophages significantly reduced as the stage of granuloma increased from stage I to stage IV (P < 0.003, P < 0.002, and P < 0.002, respectively). The distribution of immunolabeled macrophages was similar for the three markers, with immunolabeled macrophages distributed throughout early-stage granulomas (I, II), and surrounding the necrotic core in late-stage granulomas (III, IV). Our results suggest a polarization to the pro-inflammatory environment and increased expression of CD68+, iNOS+, and HLA-DR+ macrophages in the early stages of granulomas (I, II), which may play a protective role in the immune response of naturally infected beef cattle with tuberculosis.

Molecular Characterization of Cryptosporidium Oocysts in under-Five Children with Diarrhoea in Ile-Ife, Nigeria

Aim: Cryptosporidiosis is the fifth leading cause of diarrhoea-related death globally among under-five children. This study was conducted to identify and characterize oocysts of Cryptosporidium in under-five children with diarrhoea in Ile-Ife.&#x0D; Study Design: It was a case-control study.&#x0D; Place and Duration of the Study: Department of Medical Microbiology and Parasitology, Obafemi Awolowo University between April, and October 2019.&#x0D; Methodology: Ethical approval was obtained for this study. Consent was also obtained from the parents of the children. Stool samples from 53 children with diarrhoea (cases) and 94 children without diarrhoea (controls) under five years of age were collected. A structured proforma was used to obtain relevant information from the parents of the children. Stool samples were collected and examined macroscopically for consistency, appearance, and colour. Modified Ziehl-Neelsen staining was carried out on all the stool samples. The samples positive for Cryptosporidium oocysts by staining were genotyped by nested polymerase chain reaction and confirmed by sequencing of their 18S rRNA genes. Data generated were analyzed using descriptive and inferential statistics with SPSS software version 20 and STATA 15. &#x0D; Results: The prevalence of cryptosporidiosis among the study participants was 23.1% with rates of 43.4% (23) and 11.7% (11) for diarrhoeic and non-diarrhoeic children respectively. Of the 34 stool samples analysed, 6 (17.6%) were amplified by nested PCR and 4 (11.8%) were identified by sequencing. The species of Cryptosporidium identified were Cryptosporidium parvum (75%) and Cryptosporidium bovis (25%).&#x0D; Conclusion: In conclusion, Cryptosporidiosis is prevalent among under-five children with Cryptosporidium parvum and Cryptosporidium bovis as the infecting species.

Comparison of 2 PCR assays on environmental samples cultured for Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causal agent of paratuberculosis, a chronic, contagious, and incurable enteric disease of ruminants. An in-house IS900 PCR assay validated for MAP detection in sheep has been shown to have a higher sensitivity than a commercial PCR and fecal culture. We have now compared the performance of this in-house IS900 PCR assay with a commercial ISMap02 PCR assay for the detection of MAP DNA in bovine dairy farm environmental samples. We purposefully selected 30 culture-positive, 62 culture-negative, and 62 non-interpretable environmental samples. We applied the IS900 PCR assay directly to the frozen inoculum of these samples. Inocula were incubated in an automated system, and growth was confirmed by an acid-fast bacilli stain and the IS900 PCR assay. Among culture-positive samples before incubation, the IS900 PCR assay yielded significantly more positive results than the ISMap02 PCR assay; however, among culture-negative samples, the IS900 PCR assay yielded positive results both before and after incubation. The ISMap02 PCR assay did not flag positively among the culture-negative samples either before or after incubation. The IS900 PCR assay is a sensitive method that can be used to detect MAP DNA in environmental samples before incubation. The ISMap02 PCR assay is a specific method used to detect MAP DNA in environmental samples both before and after incubation.

Survey on the gastrointestinal parasites in Passeriformes and Psittaciformes with a focus on zoonotic parasites

Considering the increase in the keeping of pet birds and the importance of gastrointestinal parasitic infection, especially zoonotic parasites, this study was conducted to determine the prevalence of gastrointestinal parasites in different species of Passeriformes and Psittaciformes. In this study, one hundred and twenty fresh fecal samples were collected from different species of captive pet birds in Ahvaz from Apr 2021 to Aug 2021. The collected samples were analyzed using Clayton-Lane methods, Modified Ziehl–Neelsen, and Trichrome staining. Light microscopic morphometry was used for the identification of helminth eggs and oocysts. The results of this study showed that 37 samples (30.83%) were infected with gastrointestinal parasites. Among the helminthic parasites, three samples (2.5%) were infected with Hymenolepis spp.. Among protozoan parasites, 24 samples (20%) were infected with Eimeria spp., three samples (2.5%) were infected with Isospora spp., two samples (1.66%) were infected with Cryptosporidium spp., and five samples (4.16%) were infected with Giardia spp.. The results of the present study showed that the highest rate of gastrointestinal parasitic infection was in Passeriformes kept in pet shops and the prevalence of gastrointestinal parasitic infection was higher in the zebra finch, common mynah, and canary compared to other pet birds. But the highest rate of zoonotic parasitic infection was in Psittaciformes. According to the results, zoonotic parasites are present in pet birds in the Ahvaz area and should be considered by the owners of these pet birds, breeders, veterinarians, and public health organizations.

Aortic endograft infection by Mycobacterium abscessus subsp. massiliense with acquired clarithromycin resistance: a case report

BackgroundMycobacterium abscessus subsp. massiliense (MMA) comprises a group of non-tuberculous, rapidly growing mycobacteria. Although MMA can cause pulmonary diseases, surgical site infections, and disseminated diseases, aortic endograft infection has not been reported. Here, we describe the first case of aortic endograft infection caused by MMA.Case presentationTwo months after stent-graft insertion for an abdominal aortic aneurysm, an 85-year-old man was admitted with fever and abdominal pain and was diagnosed with aortic endograft infection. Despite 14 days of meropenem and vancomycin intravenous administration, periaortic fluid pooling increased as compared to that before antibiotic administration. The abscess was drained, and fluorescent acid-fast staining of the abscess fluid revealed bacilli. We conducted genetic tests on the genes hsp65, rpoB, and sodA, performed Whole Genome Sequencing (WGS), and identified the organism as MMA. Intravenous imipenem–cilastatin (IPM/CS), amikacin (AMK), and oral clarithromycin (CAM) were administered. After 2 months, oral CAM and sitafloxacin were administered because the abscess had decreased in size. However, after 6 weeks, the abscess increased in size again. Antimicrobial susceptibility testing of the drainage fluid from the abscess resulted in the isolation of an MMA strain that had acquired resistance to CAM. Intravenous IPM/CS, AMK, and oral linezolid were added to the treatment regimen along with oral CAM and STFX. However, he was not fully cured and died 6 months later. Neither the full-length erythromycin ribosome methyltransferase (erm)(41) gene nor the rrl or rpIV gene mutations were found by Sanger sequencing in the pre- and post-treatment strains. Whole-genome sequence analysis of the post-treatment strain revealed mutations in genes with no previous reports of association with macrolide resistance.ConclusionsAortic endograft infection caused by MMA strain is extremely rare; nonetheless, MMA should be suspected as the causative microorganism when broad-spectrum antimicrobials are ineffective.

A rare case of actinomycosis in routine fnac of the face in a tertiary care center in Central India

Actinomycosis is a chronic suppurative bacterial infection. It is caused by gram-positive, non-acid fast, filamentous, anaerobic to microaerophilic bacilli. Actinomycosis on the root of the nose is a rare entity. We report a case of 38 years old male who presented with swelling over the root of the nose since birth. Mucoid material was aspirated. Microscopic examination reveals a filamentous structured colony and crystal against the granular proteinaceous background. Features suggest benign cystic lesion and secondary actinomycosis infection, negative with an acid-fast bacilli stain consistent with Actinomyces species. As stated in most of the literature, mainly males are affected by this condition. Very little information is available on the use of fine-needle aspiration cytology (FNAC) to diagnose these lesions. We are reporting a case of an actinomycotic lesion that was reliably diagnosed with FNAC. So, we emphasized here that FNAC is aninexpensive, rapid, and simple outpatient procedure that helps in the rapid diagnosis of Actinomycosis.

Co-infection with Neisseria mucosa in a patient with tuberculous otitis media

Tuberculous otitis media (TOM) is a rare manifestation caused by Mycobacterium tuberculosis with low incidence rates among extrapulmonary tuberculosis cases. Diagnosis is often delayed because of the presence of several clinical manifestations and the high prevalence of secondary bacterial infections. Few reports have attributed secondary bacterial infections in patients with TOM to commensal Neisseria. Thus, understanding the pathogenic mechanisms and clinical features of commensal Neisseria is important, considering its recent presentation as an infection-causing pathogen. Neisseria mucosa is a commensal inhabitant in humans and is generally considered non-pathogenic but can cause infection in rare cases. Here, we report an atypical secondary infection caused by Neisseria mucosa in an 81-year-old woman with TOM being treated for pulmonary tuberculosis. Direct purulent otorrhea smear microscopy revealed no acid-fast bacilli using Ziehl-Neelsen staining, whereas the phagocytosis of gram-negative cocci by white blood cells was confirmed using Gram staining. Otorrhea culture revealed the growth of N. mucosa. Subsequently, M. tuberculosis infection in the otorrhea was identified using a culture-based method. Vigilance is critical for the early detection of TOM to prevent further complications. This report raises awareness regarding TOM and provides insight into the pathogenicity of N. mucosa in otitis media.

Health impacts of poor water quality on an endangered shorebird breeding programme in Aotearoa New Zealand

ABSTRACT Case history Two clusters of mortality among endangered tūturuatu/tchūriwat’/shore plover (Thinornis novaeseelandiae) have occurred at captive breeding facilities around New Zealand in recent years. In the first, four chicks died at Pūkaha National Wildlife Centre (Mount Bruce, NZ) in February 2016, and in the second five adult birds at the Cape Sanctuary (Cape Kidnappers, NZ) died in 2022. Clinical findings In 2016, four chicks were noted to become weak, have increased vocalisations and closed eyes prior to death. The remaining chicks were treated for 5 days with amoxycillin/clavulanate orally twice daily. Water containers and brooders were cleaned and disinfected with chlorhexidine. No further mortality was seen. In the 2022 cluster, three adult breeding birds died acutely and five others showed inappetence, weight loss and diarrhoea approximately 10 days after heavy rains flooded the local river. The five birds were treated with amoxycillin/clavulanate orally twice daily and oral fluids for 5 days. Two birds died and three survived. No breeding occurred in the aviaries in the following season. Pathological findings In 2016, the chicks showed pulmonary changes ranging from congestion and oedema to heterophilic inflammation consistent with septicaemia. In 2022, the adult birds showed proliferation of bacteria in the distal small intestine associated with mucosal ulceration and heterophilic infiltration. Acid-fast staining of the caecal contents in one bird showed organisms consistent with Cryptosporidium spp. Laboratory findings Aerobic bacterial cultures of the lung and liver of two affected chicks carried out in 2016 showed heavy growth of Plesiomonas shigelloides. The same organism was cultured from water trays and holding tanks containing water boatmen (Sigara arguta) on which the chicks were fed. In 2022, cultures from the livers of three dead birds each showed a mixed bacterial growth with differing dominant organisms (Aeromonas sobria, Hafnia alvei, Citrobacter freundii and an Enterococcus sp.). PCR and sequencing confirmed Cryptosporidium parvum in the caecum of one bird. Fresh faeces from 24 breeding birds from the captive breeding facilities were negative by PCR for Cryptosporidium spp. The captive breeding facilities obtain water for the aviaries and aquatic invertebrates to feed to the chicks from local freshwater sources. Water quality testing at the Cape Sanctuary revealed concentrations of faecal indicator bacteria in excess of safe drinking water guidelines, with peaks following heavy rainfall. Clinical relevance Fluctuations in water quality associated with mammalian faecal bacteria can adversely affect bird health and impact on captive rearing of endangered wildlife.

Value of ultrasound guided biopsy combined with Xpert Mycobacterium tuberculosis/resistance to rifampin assay in the diagnosis of chest wall tuberculosis.

The thoracic wall lesions, particularly chest wall tuberculosis, and chest wall tumors and other pyogenic wall and actinomycetes infections, almost always present as a diagnostic challenge. To explore the value of ultrasound-guided biopsy combined with the Xpert Mycobacterium tuberculosis/resistance to rifampin (MTB/RIF) assay to diagnose chest wall tuberculosis. We performed a retrospective study of patients with chest wall lesions from March 2018 to March 2021. All patients received the ultrasound-guided biopsy for pathology examination, acid-fast Bacillus staining, mycobacterial culture, and Xpert MTB/RIF analysis. The sensitivity, specificity, and area under the curve (AUC) were calculated for these diagnostic tests, either individually or combined. Rifampicin resistance results were compared between the mycobacterial culture and the Xpert MTB/RIF assay. In 31 patients with the chest wall lesion biopsy, 22 patients were diagnosed with chest wall tuberculosis. Of them, 3, 6, and 21 patients tested positive for mycobacterial culture, acid-fast stain, and Xpert MTB/RIF assay, respectively. The rifampicin resistance results of the 3 culture-positive patients were consistent with their Xpert MTB/RIF assay results. When considering the sensitivity, specificity, and AUC value, the Xpert MTB/RIF assay (95.5%, 88.9%, and 0.92, respectively) was a better choice than the acid-fast Bacillus stain (27.3%, 100.0%, and 0.64, respectively) and mycobacterial culture (13.6%, 100.0%, 0.57, respectively). No complications were reported during the procedure. Ultrasound guided biopsy combined with Xpert MTB/RIF has high value in the diagnosis of chest wall tuberculosis, and can also detect rifampicin resistance.

SAT612 Granulomatous Mastitis: An Unusual Presentation Of Prolactinoma

Abstract Disclosure: H.R. Alkaissi: None. E. Kim: None. S. McFarlane: None. Background: Granulomatous mastitis (GM) is a rare, benign inflammatory breast disease of unclear etiology with clinical presentation that is similar to inflammatory breast cancer and periductal lobular mastitis. It occurs mainly in younger females and during the postpartum period. Prolactin (PRL) is an essential hormone of pituitary and extrapituitary origin that promotes lactation and the development of mammary glands within breast tissues. In addition to its lactogenic effect, it has over 300 other effects, several of which are immunomodulatory, including the expression and secretion of proinflammatory cytokines from macrophages. Clinical Case: A 37-year-old African American woman presented with right breast pain and swelling for a few weeks. Her medical history includes amenorrhea for five years. She is gravida 1, para 1 with normal spontaneous vaginal delivery at the age of 20. Physical exam showed non-bloody, non-purulent nipple discharge bilaterally, otherwise unremarkable. A breast ultrasound showed a 1 cm complex cystic mass. Biopsy showed benign breast tissue, with noncaseating granulomas formed by macrophages (CD68 positive on immunohistochemistry). Ziehl-Neelsen staining for acid-fast bacilli was negative. Staining for fungi was also negative. Prolactin level was 237 ng/mL. TSH, IGF-1, and ACTH were within normal. Pituitary MRI showed a focus of abnormal signal and diminished enhancement measuring 2.8x3.2x4.2 mm within the left side of the pituitary gland is compatible with a microadenoma. The patient was started on bromocriptine 2.5mg daily. Discussion: Our patient represents a rare case of granulomatous mastitis in the setting of microprolactinoma, likely long-standing, given the five years history of amenorrhea. The immunomodulatory effect of PRL on several immune system cells, including B-cells, T-cells, and macrophages, might be an underlying mechanism. In a recent study of 25 samples of GM, metagenomics studies identified unusual organisms in 68% of the samples. Of these, Corynebacterium kroppenstedtii was the most common (60%), followed by Pseudomonas oleovorans (16%), Epstein-Barr virus (4%), Acinetobacter baumannii (4%), and Tepidiphilus thermophilus (4%). In addition, 50% of the patients had elevated PRL, and elevated immunological markers, such as C3 and IgA [1]. This suggests that GM results from an interaction between the presence of certain pathogens in a host with elevated PRL that mediates proinflammatory response.

Ocorrência de Cryptosporidium spp. em bezerros de rebanhos leiteiros do norte de Antioquia, Colômbia

ABSTRACT Cryptosporidium spp. are important enteropathogen protozoan parasites that infect humans and other animals throughout the world. Cryptosporidiosis in cattle industry leads to considerable economic losses due to diarrhea, dehydration, growth retardation, weight loss, and possibly death, however, data on the occurrence ofCryptosporidiumspp. in cattle in Colombia are limited. Therefore, this study aimed to investigate the occurrence and possible factors associated to the excretion of Cryptosporidium spp. oocyst in pre-weaned calves from dairy farms in Northern Antioquia, Colombia. In addition, Sheather’s sugar floatation (SSF), and Modified Ziehl-Neelsen staining (MZN) methods were compared. A total of 41 fecal samples were collected from calves between 1 and 92 days of age of which 23 were positive (56.1%). Crossbreed calves were nine times less susceptible than purebred dairy cattle to excretion of Cryptosporidium spp. oocysts (OR=0.10). MZN was the best technique for the detection of oocysts in fecal samples, however, the mean number of days to detect cryptosporidial oocysts was lower for the SSF method. Cryptosporidium is widespread among calves under 2 months of age in dairy herds of Northern Antioquia, although further investigations considering a greater number of farms and animals are necessary.

Diagnostic Efficacy of New Xpert Ultra for Extrapulmonary Tuberculosis Using Culture and Composite Reference Standard.

Xpert Ultra (Cepheid, USA) is recently introduced with an extra category of trace-positive results and higher sensitivity for tuberculosis (TB) diagnosis. The objective of the study was to assess the diagnostic accuracy of Xpert Ultra for extrapulmonary samples using culture and composite reference standard (CRS) as the gold standard. In a 1-year (March 2021-22) prospective observational study, samples of suspected extrapulmonary TB (EPTB) patients were subjected to Ziehl-Neelsen staining, culture, and Xpert Ultra (Cepheid, Sunnyvale, CA) tests. Relevant clinical and treatment information was noted. The diagnostic accuracy of Xpert Ultra compared with culture and CRS was calculated. Out of 1720 suspected patients of EPTB, 223 (13%), predominantly males 135 (60%), with a mean age of 41.46 ± 19.81 years, were diagnosed as TB positive following CRS criteria. The maximum cases were of pleural TB (35.4%), followed by central nervous system TB (17.9%), gastrointestinal TB (17.5%), and lymph node TB (12.1%). Of all samples, 150 (8.7%) were microbiologically confirmed, including 141 detected by Xpert ultra, 67 culture positive, and only 16 smear positive. Among the Xpert Ultra-positive samples, 35 showed trace results, including six false-positive results. Considering culture and CRS as the gold standard, the sensitivity (86.57%, 59.64%) and specificity (94.98%, 99.47%) of Xpert Ultra were calculated, respectively. Rifampicin resistance was detected in 1 (0.70%) sample. Diagnosis of EPTB is a challenge and Xpert Ultra may detect TB at a very early stage. However, it is essential to rule out false-positive results. Additional studies are needed on Xpert Ultra to interpret trace results better.

Single nucleotide polymorphism of IL-18 gene and resistin gene in children infection with Entamoeba histolytica

Abstract Background: Human amebiasis is caused by Entamoeba histolytica, which is found in many tropical countries. E. histolytica infections are known to have a wide range of clinical consequences. Most infections are asymptomatic; some cause diarrhea and dysentery, and only a few cause extraintestinal consequences, including liver abscess. Single nucleotide polymorphisms (SNPs) are the most prevalent kind of sequence variation in genomes and are regarded as useful genetic markers for exposing the evolutionary history and common genetic variants that explain the heritable risk for common illnesses and effected in the progression of some infections. Objective: Investigation of gene polymorphism of the parameters (interleukin 18 [IL-18], resistin) most susceptible to parasitic infection. Materials and Methods: Stool samples were collected from patients who were attending the (Kerbala Teaching Hospital for children in the holy city of Kerbala), between February 2021 and January 2022. In total, 3748 samples of feces from children between the ages of 1 and 15 years from both gender were tested using both direct smear and acid-fast stain tests in addition to rapid test techniques. Five milliliters of blood were obtained from 25 patients infected with E. histolytica, as well as 5 mL of blood from 25 healthy youngsters (sample control). Results: The result showed the distribution of genotype of IL-18 SNPs in intestinal parasites patient in contrast to the control group; there is an increase in the level of IL-18 in some SNPs, resistin concentration was statistically significantly different between resistin gene polymorphisms. Conclusion: The IL-18 and resistin polymorphism can be considered one of the genetic factors responsible for the progression of E. histolytica infection.

A Comprehensive Review and Analysis of Intestinal Parasitic Infections in School Children from South India

Background and Aim: According to the World Health Organization, close to 21% of India’s population may be affected by intestinal parasites, accounting for 39 million disability-adjusted life years, which may likely be due to substandard sanitation practices and insufficient personal hygiene standards. This research was conducted with the aim of determining the prevalence of helminthic infections among school-aged children in the South Canara district of India. Materials and Methods: We conducted a cross-sectional study involving school children from nursery to the 8th Grade in the South Canara district of Karnataka, India. The presence of various intestinal parasites in different age groups was determined by performing routine stool microscopy. Modified acid-fast staining was performed for stool samples collected from children &lt;5 years of age for the detection of intestinal coccidian parasites. Statistical analysis was carried out to analyze age-related trends in relation to the prevalence of commonly detected intestinal parasites. Results: Intestinal parasites were identified in 10 of the 253 students (3.9%), with the highest incidence observed in the age group of 7–10 years (1.9%). The most prevalent parasite identified was the cysts of Giardia lamblia, with a prevalence of 7.54%, followed by Ascaris lumbricoides and hookworm, each having a prevalence of 5.66%. Of the 253 students, 92.2% practiced handwashing with soap after defecation, while 7.8% did not use soap. The handwashing practices did not significantly vary across different age groups. Clinically, pallor was observed in 64.2% of the individuals. Conclusion: Giardiasis was the most common parasitic infection, followed by soil-transmitted helminths such as Ascariasis and hookworm. The level of hygiene practices among children exceeded the prevailing national and state standards.

P11 A case of Mycobacterium marinum infection in a patient with rheumatoid arthritis recently initiated on adalimumab

P11 A case of <i>Mycobacterium marinum</i> infection in a patient with rheumatoid arthritis recently initiated on adalimumab

Dynamics of protozoal excretion in the faeces of calves during the first 28 days after arrival at the fattening farm indicate infection before regrouping and show poor temporal correlation with diarrhoea

BackgroundCalves in dairy cattle production in Switzerland are transported to a fattening farm at the age of 3–5 weeks, and frequently suffer from diarrhoea within the first 14 days after arrival. To characterise the role of intestinal protozoa in this, we investigated the excretion dynamics of Eimeria, Cryptosporidium and Giardia during the first 28 days after the arrival and regrouping of calves at fattening farms.MethodsA total of 610 faecal samples from 122 calves (mean age 37.3 days; mean body weight 79.8 kg) were collected on seven different fattening farms during the first 28 days after the arrival and regrouping of the animals. The farms were visited between January and April (cold season; n = 4) and between June and August (warm season; n = 3). The samples were collected rectally on days 1, 4, 7, 14 and 28, assessed for consistency, and analysed using the McMaster method for quantitative determination of the number of Eimeria oocysts per gram of faeces (OPG), flotation for morphological differentiation of the unsporulated Eimeria oocysts, a concentration method for the semi-quantitative determination of Giardia cysts, and modified Ziehl–Neelsen staining for semi-quantitative determination of Cryptosporidium oocysts.ResultsOverall, 50.8% (62/122) of the animals had diarrhoea during the study period. However, the faecal excretion of protozoal pathogens was neither associated with diarrhoea nor with body weight gain of the animals. Altogether, 90.2% (110/122) of the calves were Eimeria positive. Eimeria zuernii was excreted by 51 (41.8%) and Eimeria bovis by 68 (55.7%) animals. In the warm season more animals tested positive for Eimeria and OPGs were higher than in the cold season. There was no correlation between the age of the calves and the OPG values. Overall, 64.8% (79/122) of the calves excreted Eimeria oocysts within the first 7 days, indicating that they had been infected with the parasite on the dairy farm of origin. Eighty-nine calves (73.0%) excreted Giardia cysts, with more positive animals in the cold (80.3%) compared with the warm season (64.3%). Only Giardia duodenalis assemblage E was identified. Cryptosporidium oocysts were microscopically detected in 14 animals (11.5%) on five farms. Cryptosporidium spp. were present in a total of 12 animals, i.e. Cryptosporidium parvum in nine, Cryptosporidium ryanae in two, and Cryptosporidium bovis in one animal.ConclusionsA better understanding of the temporal dynamics of protozoal infections in calves is helpful for the implementation of appropriate measures to protect the health of these animals at a critical phase in their lives. Our results indicate that factors other than those examined in the present study contributed to the onset of diarrhoea in the calves.Graphical abstract

Magnitude of tuberculosis lymphadenitis, risk factors, and rifampicin resistance at Adama city, Ethiopia: a cross-sectional study

Mycobacterium tuberculosis complex has an impact on public health and is responsible for over one million deaths per year. Substantial numbers of people infected with M. tuberculosis can develop tuberculosis lymphadenitis; however, there is a limited study in Adama, Ethiopia. The aim of this study was to determine the magnitude of Tuberculosis lymphadenitis, its predictors, and rifampicin-resistance gene-positive M. tuberculosis. A total of 291 patients with enlarged lymph nodes were recruited from May 2022 to August 30 at Adama Comprehensive Specialized Hospital Medical College (ACSHMC). GeneXpert, Ziehl–Neelsen staining, and cytology were used for the diagnosis of TB lymphadenitis from the Fine Needle Aspirate (FNA) specimen. Rifampicin-resistant gene was detected using GeneXpert. For data entry and analysis, Epi Data version 3.0 and SPSS version 25 were used respectively. A binary logistic regression model was used to identify predictors of TB lymphadenitis. A p < 0.05 with a 95% confidence interval (CI) was taken as a cut point to determine the significant association between dependent and independent variables. The prevalence of TB lymphadenitis using GeneXpert, Ziehl–Neelsen staining, and cytology were 138 (47.4%) (95% CI 41.70–53.10), 100 (34.4%) (95% CI 28.94–39.85), and 123 (42.3%) (95% CI 36.63–47.00) respectively. Nine (3.1%) participants were infected with rifampicin-resistant gene-positive M. tuberculosis. Out of the total M. tuberculosis detected by GeneXpert (n = 138), 9 (6.5%) were positive for rifampicin resistance-gene. Participants with a chronic cough had 2 times odds of developing TB lymphadenitis (AOR: 2.001, 95% CI 1.142–3.508). Close to half of patients with enlarged lymph nodes were positive for M. tuberculosis by the GeneXpert method in the study area. Chronic cough was significantly associated with TB lymphadenitis. Rifampicin-resistant gene-positive M. tuberculosis was relatively prevalent among patients with enlarged lymph node in the study area.

EBUS-GS with the GeneXpert MTB/RIF assay for diagnosis of Mycobacterium tuberculosis infection of isolated pulmonary nodules

ObjectiveInvestigate the use of endobronchial ultrasonography with a guide sheath (EBUS-GS) combined with Gene Xpert MTB/RIF (Xpert) for diagnosis of Mycobacterium tuberculosis (MTB) infection in isolated pulmonary nodules.MethodsPatients who had isolated pulmonary nodules and unknown diagnoses at our institution from October 2020 to December 2021 were prospectively examined using EBUS-GS and Xpert. The diagnostic values of using EBUS-GS or bronchoalveolar lavage fluid (BALF) with acid-fast staining, MGIT 960 culture, pathological examination, and Xpert for isolated pulmonary nodules caused by MTB infection were compared using receiver operating characteristic (ROC) analysis.ResultsThere were 135 patients, 64 with isolated pulmonary tuberculomas and 71 with non-tuberculous lesions. The sensitivity of EBUS-GS with Xpert was significantly higher than BALF with Xpert (57.81% vs. 34.78%, P = 0.017). Use of EBUS-GS with Xpert and MGIT 960 culture further increased the sensitivity to 62.50% (95%CI 50.64–74.36) and increased the specificity to 100%. The AUC values of BALF with MGIT 960 culture was 0.663(95%CI 0.543–0.783) and BALF with Xpert was 0.674 (95%CI 0.556–0.792). The AUC values of EBUS-GS with MGIT 960 culture was 0.680 (95%CI 0.554–0.743), with pathological examination was 0.713 (95%CI 0.573–0.760), and with Xpert was 0.789 (95%CI 0.655–0.829).ConclusionUse of EBUS-GS with Xpert had high sensitivity and specificity in the diagnosis of isolated pulmonary tuberculoma. This method has significant potential for use in clinical practice.

A Retrospective Analysis of the Clinicopathological Features of Patients with Tuberculous Mastitis: A Single Institution’s Experience

Introduction: Tuberculosis remains a global health burden, especially in low- and middle-income countries. Breast tuberculosis is a rare disease with minimal research available. This disease produces a diagnostic challenge as the clinical presentation is variable, and diagnosis often requires additional investigations. This study was undertaken to determine the efficacy of cytology and histology, together with ancillary studies, in diagnosing tuberculous mastitis. Methods: This retrospective study was conducted in a Johannesburg Hospital over 5 years. Thirty-two patients with confirmed tuberculous mastitis were included. The patients were considered positive for tuberculous mastitis if histological or cytological findings were confirmed with either a positive tuberculosis culture, Ziehl-Neelsen stain, or polymerase chain reaction examination/GeneXpert. Results: This case series comprises 3 males and 29 females with a mean age of 35.66. A breast mass was the most common presentation. Over these 5 years, more biopsies were performed on inflammatory breast lesions than fine needle aspirations. There was a higher confirmation rate for cytology diagnoses compared to histology diagnoses. Conclusion: This study supports using fine needle aspiration combined with GeneXpert as the primary diagnostic modality in diagnosing tuberculous mastitis. This test combination is advantageous in resource- and financially constrained environments as it is relatively simple to perform, cost-effective, and has a rapid turnaround time.