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Background & objectives: Surveillance of canine leishmaniasis in Colombia is restricted to the appearance of visceral leishmaniasis cases in humans, and is mainly performed by serological tests. This requires blood sampling by veterinarians or technicians according to Colombian laws. The aim of this study was to evaluate the utility of conjunctival swabs in the molecular detection of Leishmania in dogs from the municipality of Ovejas, Sucre. Methods: The present study was cross-sectional and descriptive. The collection source of samples and information was primary. Blood samples and conjunctival swabs from 121 dogs were analysed by PCR-ITS1 to detect Leishmania spp. Positive samples were used to amplify a conserved region of the Leishmania infantum kinetoplast minicircle. Performance of both sample types was calculated by proportion of positive samples of each type and the degree of agreement between them was determined by Cohen’s kappa (κ) agreement index. Results: Leishmania infection was detected in 17.4% (21/121) of blood samples and in 16.5% (20/121) of conjunctival swabs. In total, 28.1% (34/121) of the canines were infected, of which 11.8% (4/34) were infected with L. infantum in the conjunctival swabs and 5.9 % (2/34) in the blood samples. The agreement between blood and conjunctiva was medium (κ = 0.207) by PCR-ITS1 amplification. Interpretation & conclusion: The use of conjunctival swab as a non-invasive sample could be used as an alternative method for surveillance of canine leishmaniasis.

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Objective: To evaluate the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the tear and conjunctival secretions of laboratory-confirmed coronavirus disease-2019 (COVID-19) patients. Methods: A total of 152 consecutive COVID-19 patients, confirmed by real-time polymerase chain reaction (RT-PCR) test in nasopharyngeal swabs, were included. The conjunctival swabs were taken from both eyes by the same ophthalmologist in the first 24-36 hours after positive test results for COVID-19 in the nasopharyngeal swabs were detected. Results: Of the 152 patients, 96 (63.15%) were male and 56 (36.85%) were female. The mean age was 39.36±13.15 years. Sixteen (10.5%) patients had symptoms or findings such as tearing, stinging, redness, pain, and burning in their eyes. RT-PCR tests for SARS-CoV-2 RNA in conjunctival swabs were positive in 13 (8.55%) of 152 COVID-19 patients. Conclusion: This study showed that SARS-CoV-2 may be found in the conjunctival swabs of laboratory-confirmed COVID-19 patients, and ocular secretions may be a possible route for virus transmission. [ABSTRACT FROM AUTHOR]

Purpose: To detect the viral RNA load of SARS-CoV-2 in conjunctival swabs of COVID-19 patients, and compare with nasopharyngeal swabs. Methods: Conjunctival swabs of COVID-19 patients (with PCR positive nasopharyngeal swabs) were subjected to quantitative reverse transcription–polymerase chain reaction (RT-PCR) for detection of SARS-CoV-2 RNA. The cycle threshold (Ct) values of Open Reading Frame 1 (ORF 1 Ab gene) and nucleoprotein (N gene) PCRs were used to assess the viral RNA load, and compare them with the baseline values of nasopharyngeal swabs. Results: Of 93 patients, 17 (18.27%) demonstrated SARS-CoV-2 RNA in conjunctival swabs. Baseline nasopharyngeal swabs were collected at a median of 2 days; while, the conjunctival swabs were collected at median 7 days, from onset of illness (p < 0.001). Despite a significant delay in conjunctival swab collection than nasopharyngeal swabs, the Ct values (ORF or N gene PCRs) were comparable between nasopharyngeal swab and conjunctival swab samples. Subsequently, during the recovery period, in four of these 17 patients (with conjunctival swab positivity), when the second nasopharyngeal swab was ‘negative’, the conjunctival swab was ‘positive’. Conclusion: The conjunctival swabs demonstrated SARS-CoV-2 RNA in 17 (18.27%) of 93 COVID-19 patients. Our results may suggest a delayed or a prolonged shedding of the virus/viral RNA on the ocular surface than in nasopharyngeal mucosa. [ABSTRACT FROM AUTHOR]

IntroductionPurulent conjunctival discharge in hospitalized preterm infants may indicate conjunctivitis and warrant treatment. The purpose of this study was to examine the relationship between positive conjunctival swab (CS) culture and late-onset sepsis (LOS) in preterm infants.MethodsA retrospective cohort study was conducted to determine the relationship between positive CS culture growth results (CSP) obtained in preterm infants ≤34 weeks' gestation and the development of LOS within 120 h of obtaining CS compared with those who had negative CS culture results (CSN). Electronic medical records were reviewed from January 2015 until December 2019 for preterm infants presenting with purulent conjunctival discharge and underwent CS culture testing due to suspected conjunctivitis.ResultsOf the 234 CS cultures obtained during the study period, 145 (61.9%) were CSP compared to 89 (38.1%) CSN cultures. Gram-negative organisms accounted for 70% of all CSP cultures, with the remaining 30% being Gram-positive. Patients with CSP were smaller, younger, had lower 1-minute APGAR scores, and required respiratory support more frequently than those with CSN. Infants with CSP received antibiotics for longer periods, both topically and systemically. Infants who developed LOS were more likely to require invasive ventilation (adjusted odds ratio, 33.5; 95% CI, 2.52–446.5, p = 0.008). The incidence of LOS between the two groups was similar, with 6.2% observed in the CSP group compared to 3.4% in the CSN group (p = 0.543). Similarly, the rates of bacteremia were similar in both groups. Of the CSP patients who were presented with bacteremia, four out of seven (57%) exhibited bacteremia caused by the same organism found in their CS cultures. Similarly, within the entire cohort, respiratory cultures were performed on nine intubated patients within two weeks of obtaining CS cultures. Of these, in the CSP group, five out of six (83%) showed an organism identical to that found in the CS cultures.ConclusionThe study found a significant proportion of positive CS cultures in preterm infants, with distinct patient characteristics and treatment compared to negative cultures. While the incidence of LOS was not significantly different between the two groups, some CSP patients demonstrated bacteremia with the same CS organism, suggesting a possible connection between conjunctival or respiratory colonization and bacteremia.

Background: The composition of the normal human ocular surface flora includes Staphylococcus epidermidis, Staphylococcus aureus, and diphtheroids. There are several factors that may alter the composition of ocular surface flora: age, dry eye, immunosuppressive medication and medical illnesses such as diabetes mellitus. Patients in the intensive care unit (ICU) frequently have impaired ocular defence mechanisms as a result of metabolic derangements, mechanical ventilation, sedation, paralysis and decreased level of consciousness. Dry eye is one of the major risk factors of infectious keratitis. Intensive care unit (ICU) patients are susceptible to a number of ophthalmic conditions that may result in visual loss. Aims and Objectives: To evaluate the Conjunctival Swab Culture among Patients admitted to the Intensive Care Unit who are on ventilators. Materials and Methods: Inclusion Criteria :1. All patients aged 18-65 years admitted to RICU for a time period >48 hours were included. 2. Neonates in NICU. Exclusion criteria: Patients who presented with ocular surface disorders prior to admission in RICU. Results: In this study, a total of 28 patients were taken. Among these are 16 males, 4 females and 8 neonates. In this study, 8 cases positive for Gram-positive cocci were seen in singles, pairs, tetrads and clusters in the right eye and 7 cases in the left eye. Coagulase-negative staphylococci were isolated in 6 cases in the right eye and 4 cases in the left eye. Conclusion: ICU medical and nursing staff are primarily concerned with life-threatening conditions; therefore, the ocular signs and symptoms may be missed, leading to serious ocular complications, including corneal ulceration and infectious keratitis. For these reasons, meticulous eye care with regular cleaning of the eyes, installation of lubricating drops and ointments, and consultation from an ophthalmologist in case of a suspected infection are recommended. [ABSTRACT FROM AUTHOR]

Abstract Objectives This study aimed to investigate the spectrum of ocular characteristics and viral presence in the conjunctival swab of patients with COVID-19. Methods In this cross-sectional study, fifty-three patients were recruited from two COVID-19 referral hospitals in Jakarta (Cipto Mangunkusumo Hospital and Persahabatan Hospital) from July 2020 to March 2021. The inclusion criteria were patients who were suspected of or confirmed cases of COVID-19 with or without ocular symptoms. Demographic data, history of COVID-19 exposure, underlying medical condition, systemic symptoms, ocular symptoms, supporting laboratory results, reverse-transcriptase polymerase chain reaction (RT-PCR) of naso-oropharyngeal (NOP) swab and conjunctival swab were collected. Results Fifty-three patients who were suspected, probable or confirmed cases of Covid-19 were included. Forty-six out of 53 patients (86.79%) tested positive for either Covid-19 antibody rapid test or naso-oropharyngeal (NOP) swab. Forty-two patients tested positive for NOP swab. Fourteen out of 42 patients (33.33%) experienced symptoms of ocular infection including red eye, epiphora, itchy eyes, and eye discharge. None of these patients were tested positive for conjunctival swab. Two out of 42 patients (4.76%), who were tested positive for conjunctival swab, did not experience any ocular symptoms. Conclusions Establishing the relationship between Covid-19 infection, ocular symptoms, and presence of SARS-CoV-2 virus on the ocular surface proves to be challenging. In Covid-19 patients, ocular symptoms did not warrant a positive conjunctival swab result. On the contrary, a patient without ocular symptoms can also have detectable presence of SARS-CoV-2 virus on the ocular surface.

Objectives: This study aimed to investigate the spectrum of ocular characteristics and viral presence in the conjunctival swab of patients with COVID-19. Methods: In this cross-sectional study, fifty-three patients were recruited from two COVID-19 referral hospitals in Jakarta (Cipto Mangunkusumo Hospital and Persahabatan Hospital) from July 2020 to March 2021. The inclusion criteria were patients who were suspected of or confirmed cases of COVID-19 with or without ocular symptoms. Demographic data, history of COVID-19 exposure, underlying medical condition, systemic symptoms, ocular symptoms, supporting laboratory results, reverse-transcriptase polymerase chain reaction (RT-PCR) of naso-oropharyngeal (NOP) swab and conjunctival swab were collected. Results: Fifty-three patients who were suspected, probable or confirmed cases of Covid-19 were included. Forty-six out of 53 patients (86.79%) tested positive for either Covid-19 antibody rapid test or naso-oropharyngeal (NOP) swab. Forty-two patients tested positive for NOP swab. Fourteen out of 42 patients (33.33%) experienced symptoms of ocular infection including red eye, epiphora, itchy eyes, and eye discharge. None of these patients were tested positive for conjunctival swab. Two out of 42 patients (4.76%), who were tested positive for conjunctival swab, did not experience any ocular symptoms. Conclusions: Establishing the relationship between Covid-19 infection, ocular symptoms, and presence of SARS-CoV-2 virus on the ocular surface proves to be challenging. In Covid-19 patients, ocular symptoms did not warrant a positive conjunctival swab result. On the contrary, a patient without ocular symptoms can also have detectable presence of SARS-CoV-2 virus on the ocular surface. [ABSTRACT FROM AUTHOR]

Purpose: To assess the presence of viral RNA in conjunctival secretions and tears of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected patients in terms of a possible ocular transmission route and also to examine whether a difference between the patients with and without ocular surface positivity existed. Methods: A prospective cross-sectional study of 70 consecutive patients who were hospitalized in Haseki Training and Research Hospital with SARS-CoV-2 from February 1 to April 1, 2021, was performed. Tears and conjunctival secretions were collected within 24 h of nasopharyngeal sample collection and examined for SARS-CoV-2 using reverse transcription real-time polymerase chain reaction (qPCR). The clinical data, results of blood tests and nasopharyngeal and conjunctival swabs, and CT findings were evaluated for all patients. Results: Seventy patients (37 males, 33 females) were included in this study. Tear–conjunctival samples from eight patients (11.42%) yielded positive PCR results although these eight patients had no eye symptoms or conjunctivitis. In patients with positive conjunctival PCR results, cycle threshold values for conjunctival samples were higher than those for nasopharyngeal samples. All findings (except gender) were similar between patients with either positive or negative conjunctival swab samples. All patients with positive conjunctival swab samples were male; however, the male ratio in patients with negative conjunctival swab samples was only 46.77%. Conclusion: In our study, the rate of conjunctival swab PCR positivity was 11.42%. It appears that even in the absence of ocular symptoms, SARS-CoV-2 virus may be present on the ocular surface; therefore, the ocular surface may be a significant viral transmission route. [ABSTRACT FROM AUTHOR]

Aims and objectives: Purpose of this study is to detect the presence of SAR-CoV-2 viral RNA in conjunctival secretions of COVID-19 patients and to compare the RT-PCR positivity rate for SARS-CoV-2 in conjunctival and nasopharyngeal swab. Materials and method: Eighty hospitalised COVID-19 patients whose nasopharyngeal swab tested positive for SARS-CoV-2 by RT-PCR were included in the study. Conjunctival swab was collected from eyes of these patients and sent for detection of SARS-CoV-2 by RT-PCR method. Results: Among the eighty patients, 51 (63.7%) were males and 29 (36.3%) were females. The mean age of the patients was 55.93 ± 16.59. Six patients had ocular manifestations. Eleven (13.75%) patients tested positive on conjunctival swab for SARS-CoV-2 viral RNA and only one of them had ocular manifestations out of the eleven. Conclusion: In our study the presence of SARS-CoV-2 in conjunctival secretions of COVID-19 patients was detected and this was not dependent on presence of ocular manifestations or duration of disease. Though the conjunctival positivity is lower compared to the nasopharyngeal swab sampling, ocular surface and secretions can be a potential route of viral transmission.

Aims and objectives: Purpose of this study is to detect the presence of SAR-CoV-2 viral RNA in conjunctival secretions of COVID-19 patients and to compare the RT-PCR positivity rate for SARS-CoV-2 in conjunctival and nasopharyngeal swab. Materials and method: Eighty hospitalised COVID-19 patients whose nasopharyngeal swab tested positive for SARS-CoV-2 by RT-PCR were included in the study. Conjunctival swab was collected from eyes of these patients and sent for detection of SARS-CoV-2 by RT-PCR method. Results: Among the eighty patients, 51 (63.7%) were males and 29 (36.3%) were females. The mean age of the patients was 55.93 ± 16.59. Six patients had ocular manifestations. Eleven (13.75%) patients tested positive on conjunctival swab for SARS-CoV-2 viral RNA and only one of them had ocular manifestations out of the eleven. Conclusion: In our study the presence of SARS-CoV-2 in conjunctival secretions of COVID-19 patients was detected and this was not dependent on presence of ocular manifestations or duration of disease. Though the conjunctival positivity is lower compared to the nasopharyngeal swab sampling, ocular surface and secretions can be a potential route of viral transmission.

Abstract Background The aim of this study was to demonstrate the presence of the virus in tear and conjunctival secretions of clinically-confirmed COVID-19 pneumonia patients. Methods This prospective study was conducted at Bakirkoy Dr. Sadi Konuk Training and Research Hospital (2020/190). Nasopharyngeal and ocular samples were obtained by swab technique and investigated by RT-PCR. Results A total of 83 patients were included. The mean age was 61.88 ± 16.04 years. 28.92% of the patients had mild, 65.06% moderate and 6.02% severe pneumonia radiologically. RT-PCR was positive in 31 (37.35%) patients in the first nasopharyngeal swabs and in 19 (22.89%) in the second swabs. 17 of 19 patients had positive both first and second nasopharyngeal swabs; only the second swabs of two patients were positive. The first conjunctival swabs RT-PCR were positive in 5 out of 83 clinically-confirmed patients or 33 laboratory-confirmed patients (rates: 6.02% and 15.15%). There were no positives detected in the second conjunctival swabs. Conclusions SARS-CoV-2 can be detected in the conjunctival swabs of patients with COVID-19 pneumonia.

Manal Hadrawi,1 Mohammad Malak,1 Faeeqah Almahmoudi,1 Ahmed Mogharbel,2 Omar Rozy,1 Somaya Hanafi,1 Batool Ali,3 Sanaa Nabeel,3 Fatimah Faqieha,4 khaled Alzahrani1 1Department of Ophthalmology, King Fahad Armed Forces Hospital, Jeddah, Saudi Arabia; 2Faculty of Medicine, King Abdulalziz University, Jeddah, Saudi Arabia; 3Department of Infectious Diseases, East Jeddah Hospital, Ministry of Health, Jeddah, Saudi Arabia; 4Department of Public Health, Ministry of Health, Jeddah, Saudi ArabiaCorrespondence: khaled Alzahrani; Ahmed MogharbelDepartment of Ophthalmology, Faculty of Medicine, King Fahad Armed Forces Hospital King Abdulaziz University, Jeddah, Saudi ArabiaEmail khaledod@gmail.com; ahmadmogharbel@gmail.comObjective: To investigate the incidence of COVID-19 viral RNA in the conjunctiva of patients priorly confirmed to have COVID-19, using a conjunctival swab and to determine swab sensitivity.Methods: This cross-sectional study enrolled patients who tested positive for COVID-19 with PCR via nasopharyngeal swabs. Conjunctival swabs were collected for each patient and subjected to RT-PCR assay. Demographic data and clinical history of patients were investigated and analysed.Results: In this study, 4.9% of the patients tested positive with conjunctival swabs for COVID-19; 29.9%, 28.7%, 20.1%, and 12.2% of the patients had fever, shortness of breath, cough, and red eye, respectively. Among all patients, 18.9% and 73.2% had a history of traveling and contact with positive COVID-19 cases, respectively. There were significant correlations between the conjunctival results and diabetes (P=0.049) and hypertension (P=0.002). Traveling was a risk factor for positive conjunctival swab testing (P=0.016). The sensitivity of the conjunctival swab was 4.8%.Conclusion: Among all positive cases of COVID-19, a small percentage had positive results when tested using a conjunctival swab. Conjunctival swabs have very low sensitivity for the detection of COVID-19. However, tears could still be a mode of disease transmission, especially from patients to eye care specialists.Keywords: COVID-19, conjunctival swab, sensitivity, specificity, RNA

The novel coronavirus infection (COVID-19) caused by SARS-CоV-2 virus, may lead to ocular diseases besides affecting the respiratory system. Cororonavirus infection may be transmitted not only through the respiratory system but also through the conjunctiva. The review article summarizes the data on the origin and variants of coronaviruses that infect humans, as well as on SARS-CоV-2 structures. Literature data on prospects of molecular diagnostics of the disease, coronavirus type detection methods, clinical sample types and duration of the disease before it is diagnosed are discussed. The data on the relationship between the coronavirus infection and conjunctivitis are given. Molecular tests of the tear and conjunctival swabs were used to diagnose the coronavirus infection independently of whether symptoms of ocular disorders were present. The real-time reverse-transcription of RNA polymerase chain reaction (PCR) was the most informative method for diagnosis of SARS-CоV-2 in the early stage of COVID-19 (until the 9th day). The presence of SARS-CоV-2 in the tear fluid and conjunctival swabs indicates the need for safety measures to prevent virus transmission through the ocular surface, including protective goggles to be worn by healthcare workers. Since side effects in the eye may appear, COVID-19 patients with visual impairment who received etiotropic or pathogenetic treatment are advised to consult an ophthalmologist.

Purpose: To evaluate the presence of SARS-CoV-2 RNA in the conjunctival swab sample of positive confirmed COVID-19 patients and to find out its association with comorbidity and severity of COVID-19 disease. Methods: We conducted an observational cross-sectional study at a dedicated tertiary COVID-19 hospital in central India for a period of 8 weeks from February 2021to March 2021. We included patients who tested positive for SARS-CoV-2 RNA through nasopharyngeal swab and were above 18 years of age. Swab samples have been collected within 48 h of admission. Conjunctival swab was taken from the lower fornix of both eyes and sent to microbiology laboratory for real-time- polymerase chain reaction (RT-PCR). Results: Out of 150 patients, conjunctival swab RT-PCR was positive in five patients (3.33%). Two patients had conjunctival manifestations in the form of conjunctivitis but conjunctival swab RT-PCR was negative in those patients. Among the RT-PCR positive patients, two (40%) were from mild, one (20%) was from moderate, and two (40%) were from severe category. No association could be established between conjunctival swab RT-PCR positivity and severity of the disease or associated comorbidity. Conclusion: Our study provides evidence that SARS-CoV-2 RNA could be detected in conjunctival secretions, and though the risk is relatively low, the eye may act as source of transmission. Extra caution should be taken by healthcare workers, and use of proper precautions like face shields and goggles should be encouraged.

Objective: To determine whether tropicamide, fluorescein, and proparacaine applied topically before sample collection affect the quantity or species of bacteria isolated via aerobic culture. Animals studied: 12 female adult research beagle cross‐breed dogs. Procedures: A conjunctival swab was taken before and after the sequential application of proparacaine, tropicamide, and fluorescein to the same eye (P/T/F) with a five‐minute gap between medications. Paired swabs were submitted for aerobic culture. Bacterial enumeration was performed using the spread plate method. Following a one‐week washout period, the procedure was repeated using balanced salt solution (BSS). Following a second one‐week washout period, the experiment was repeated using ofloxacin 0.3% solution. Colony counts were compared using one‐way ANOVA and Tukey post hoc comparison. Bacterial species reduction was compared using a Friedman rank test and Dunn's method. Results: The bacterial colony count for P/T/F and BSS was significantly higher than the ofloxacin group (p = 0.0052, p = 0.0022). There was no significant difference for colony counts between P/T/F and BSS (p = 0.9295). The most frequently isolated bacteria included: Psychrobacter spp., Staphylococcus spp., Corynebacterium spp., and Streptococcus spp. The bacterial species reduction for P/T/F and BSS was significantly lower than for ofloxacin (p < 0.0001, p = 0.0160). There was no significant difference for species reduction between P/T/F and BSS (p = 0.3749). Conclusions: The application of proparacaine, tropicamide, and fluorescein did not significantly decrease the amount or species of bacteria isolated from the conjunctiva in this canine population. The application of these solutions prior to ocular swab collection in healthy dogs is unlikely to affect subsequent culture results. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the presence of SARS-CoV-2 RNA in the conjunctival swab sample of positive confirmed COVID-19 patients and to find out its association with comorbidity and severity of COVID-19 disease. Methods: We conducted an observational cross-sectional study at a dedicated tertiary COVID-19 hospital in central India for a period of 8 weeks from February 2021to March 2021. We included patients who tested positive for SARS-CoV-2 RNA through nasopharyngeal swab and were above 18 years of age. Swab samples have been collected within 48 h of admission. Conjunctival swab was taken from the lower fornix of both eyes and sent to microbiology laboratory for real-time- polymerase chain reaction (RT-PCR). Results: Out of 150 patients, conjunctival swab RT-PCR was positive in five patients (3.33%). Two patients had conjunctival manifestations in the form of conjunctivitis but conjunctival swab RT-PCR was negative in those patients. Among the RT-PCR positive patients, two (40%) were from mild, one (20%) was from moderate, and two (40%) were from severe category. No association could be established between conjunctival swab RT-PCR positivity and severity of the disease or associated comorbidity. Conclusion: Our study provides evidence that SARS-CoV-2 RNA could be detected in conjunctival secretions, and though the risk is relatively low, the eye may act as source of transmission. Extra caution should be taken by healthcare workers, and use of proper precautions like face shields and goggles should be encouraged. [ABSTRACT FROM AUTHOR]

Introduction SARS-CoV-2 (severe acute respiratory syndrome-coronavirus-2) is a coronavirus that causes COVID-19 (coronavirus disease 2019) with mild to severe respiratory illness. It is a highly contagious disease transmitted through direct or indirect contact with infected people or contaminated surfaces, mainly through respiratory droplets, but other routes are being investigated. Objective It is known that coronaviruses (CoVs) can cause a variety of ocular pathologies in animals, including conjunctivitis, anterior uveitis, retinitis, and optic neuritis, many of which are severe. However, there is no evidence of the SARS-CoV-2 presence in the eye tissue of asymptomatic patients, even if the symptomatic incidence is low. This systematic review presents updated literature on this issue. Abbreviated description of the state of knowledge COVID-19 has now spread throughout the continents and poses a global threat to public health. The risk of rapidly overloading health care systems and causing substantial mortality worldwide is real. On 11 March 2020, the World Health Organization (WHO) announced coronavirus as a global pandemic. Several studies described a few cases with initial ocular symptoms followed by systemic symptoms of the disease. Summary Although the frequency of transmission of SARS-CoV-2 infection through the eye is low, ocular symptoms are not uncommon in COVID-19. In some cases, eye symptoms may be the first signs of illness. This implies the need for hygienic recommendations and use of personal protective equipment (PPE) for medical staff and other services to minimize COVID-19 infection of both health-care workers and patients. A triage for ophthalmic outpatient clinic is mandatory.

Background: The aim of this study was to demonstrate the presence of the virus in tear and conjunctival secretions of clinically-confirmed COVID-19 pneumonia patients.Methods: This prospective study was conducted at Bakirkoy Dr. Sadi Konuk Training and Research Hospital (2020/190). Nasopharyngeal and ocular samples were obtained by swab technique and investigated by RT-PCR.Results: A total of 83 patients were included. The mean age was 61.88 ± 16.04 years. 28.92% of the patients had mild, 65.06% moderate and 6.02% severe pneumonia radiologically. RT-PCR was positive in 31 (37.35%) patients in the first nasopharyngeal swabs and in 19 (22.89%) in the second swabs. 17 of 19 patients had positive both first and second nasopharyngeal swabs; only the second swabs of two patients were positive. The first conjunctival swabs RT-PCR were positive in 5 out of 83 clinically-confirmed patients or 33 laboratory-confirmed patients (rates: 6.02% and 15.15%). There were no positives detected in the second conjunctival swabs.Conclusions: SARS-CoV-2 can be detected in the conjunctival swabs of patients with COVID-19 pneumonia. [ABSTRACT FROM AUTHOR]

The common approach of the diagnosis of Alzheimer's Disease (AD) is made with an analysis of the cerebrospinal fluid or the study of retinal fundus and the plaques formation through optical corneal tomography (OCT), or more simply with a fundus camera. Tears analysis is widely discussed in literature as an essential method to describe molecular and biochemical alterations in different diseases. The aim of our study was the identification with immunocytochemistry of Amyloid Beta-42 in tears from patients with or without familiarity for Alzheimer Disease, in order to make the diagnosis earlier and more accessible compared to other invasive methods. Our study was performed on tears from three phenotypically healthy subjects: two of them were Caucasian with Alzheimer familiarity (48 and 55 years old) and the other one was Asian without Alzheimer familiarity (45 years old) and affected by an adenoviral keratoconjunctivitis at the moment of withdrawal. Tear samples were collected from eye fornix and were examinated by immunocytochemistry (ICC) assay using anti-Amyloid Beta X-42 antibody. Two out of three tears samples showed positive Amyloid Beta-42. Considering that our patients were phenotypically healthy, the identification of Amyloid Beta-42 by ICC could be a candidable method to make the diagnosis of the disease earlier and more accessible and available then other current and invasive methods and it could be a candidate for a screening method too. [ABSTRACT FROM AUTHOR]

ObjectivesTo determine the role of equine herpesvirus (EHV) in idiopathic keratoconjunctivitis in horses and to determine whether sample collection method affects detection of EHV DNA by quantitative polymerase chain reaction (qPCR).Animals studiedTwelve horses with idiopathic keratoconjunctivitis and six horses without signs of ophthalmic disease.ProceduresConjunctival swabs, corneal scrapings, and conjunctival biopsies were collected from 18 horses: 12 clinical cases with idiopathic keratoconjunctivitis and six euthanized controls. In horses with both eyes involved, the samples were taken from the eye judged to be more severely affected. Samples were tested with qPCR for EHV-1, EHV-2, EHV-4, and EHV-5 DNA. Quantity of EHV DNA and viral replicative activity were compared between the two populations and among the different sampling techniques; relative sensitivities of the sampling techniques were determined.ResultsPrevalence of EHV DNA as assessed by qPCR did not differ significantly between control horses and those with idiopathic keratoconjunctivitis. Sampling by conjunctival swab was more likely to yield viral DNA as assessed by qPCR than was conjunctival biopsy. EHV-1 and EHV-4 DNA were not detected in either normal or IKC-affected horses; EHV-2 DNA was detected in two of 12 affected horses but not in normal horses. EHV-5 DNA was commonly found in ophthalmically normal horses and horses with idiopathic keratoconjunctivitis.ConclusionsBecause EHV-5 DNA was commonly found in control horses and in horses with idiopathic keratoconjunctivitis, qPCR was not useful for the etiological diagnosis of equine keratoconjunctivitis. Conjunctival swabs were significantly better at obtaining viral DNA samples than conjunctival biopsy in horses in which EHV-5 DNA was found.

The aim is to comparatively evaluate the results of simultaneous conjunctiva and oropharynx–nasopharynx (ONP) swabs in patients who had presented to the outpatient department with a suspicion of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2). An ONP sample was obtained following bilateral conjunctiva swabs in 85 subjects with a contact history or symptoms but unknown SARS‐CoV‐2 status and with no ocular symptoms or findings. The results were evaluated according to the patient's symptoms and how the swab was taken. The conjunctiva swab was positive in 29 (34.1%) cases and the ONP swab in 20 (23.5%) cases. Both methods produced positive results in 11 (14.1%) cases. The mean cycle threshold (Ct) value was 30.15 ± 3.41 in symptomatic cases and 33.62 ± 1.76 in asymptomatic cases (p =.008). The mean Ct value was 24.37 ± 3.48 when only the ONP swab was positive and 31.22 ± 1.99 when only the conjunctiva swab was positive. In cases that were positive by both methods, the mean Ct value was 25.21 ± 4.94 for the ONP swab and 30.29 ± 5.05 for the conjunctiva swab. We found higher SARS‐CoV‐2 detection rates with the conjunctiva swab than the ONP swab in cases with unknown SARS‐CoV‐2 status in the early period. In addition, the conjunctival viral load seemed to be higher in symptomatic cases than in asymptomatic cases. We, therefore, believe a conjunctiva swab could be an alternative method to detect SARS‐CoV‐2 at the time of the first presentation to the outpatient department. [ABSTRACT FROM AUTHOR]

Purpose: To detect the presence of viral RNA of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in conjunctival swab specimens of coronavirus disease-19 (COVID-19) patients. Methods: Forty-five COVID-19 patients positive for real-time reverse transcription-polymerase chain reaction (RT-PCR) for SARS-CoV-2 in nasopharyngeal swab with or without ocular manifestations were included in the study. The conjunctival swab of each patient was collected by an ophthalmologist posted for COVID duty. Results: Out of 45 patients, 35 (77.77%) were males and the rest were females. The mean age was 31.26 ± 12.81 years. None of the patients had any ocular manifestations. One (2.23%) out of 45 patients was positive for RT-PCR SARS-CoV-2 in the conjunctival swab. Conclusion: This study shows that SARS-CoV-2 can be detected in conjunctival swabs of confirmed cases of COVID-19 patients. Though the positivity rate of detecting SARS-CoV-2 in conjunctival swabs is very less, care should be exercised during the ocular examination of patients of COVID-19.

To investigate the presence of SARS-CoV-2 RNA in tears of patients with moderate to severe coronavirus disease 2019 (COVID-19). Cross-sectional study. Patients with laboratory-proven moderate to severe COVID-19. Tears were collected within 48 hours of laboratory confirmation using 3 methods: conjunctival swab plus Schirmer's test strips (group 1), conjunctival swab (group 2), and Schirmer's test strips (group 3). Samples from both the eyes of each patient were transported in a single viral transport media for real-time RT-PCR. Detailed demographic profiles, systemic symptoms, comorbidities, and ocular manifestations were noted. Viral load of a sample was determined using cycle threshold (Ct) value of E gene. A specimen was considered to show positive results if the amplification curve for the E gene crossed the threshold line within 35 cycles and if it showed positive results on an RNA-dependent RNA polymerase or open reading frame 1b gene assay. Of the 78 patients enrolled in the study, samples from 3 patients were found to be inadequate for analysis. Thirty-six patients (48%) had moderate disease, whereas 39 patients (52%) had severe disease, with no ocular involvement in any patient. In the 75 patients, RT-PCR analysis of tears showed positive results in 18 patients (24%), and 29 of 225 samples (12.9%) showed positive results. Positive results were found in 11 (14.7%), 11 (14.7%), and 7 (9.3%) patients in groups 1, 2, and 3, respectively (P = 0.3105). Mean Ct values in groups 1, 2, and 3 were 28.36 ± 6.15, 29.00 ± 5.58, and 27.86 ± 6.46 (P = 0.92), respectively. Five patients showed positive RT-PCR results by all 3 methods (mean Ct value, 25.24 ± 6.33), and 12 patients showed positive results by any of the 3 methods (mean Ct value, 32.16 ± 1.94), the difference in Ct values being statistically significant (P = 0.029). The median value of symptomatology in patients with positive RT-PCR results from tears was 5 days (range, 4–9 days). SARS-CoV-2 RNA was detected in tears of 24% of patients with laboratory-proven moderate to severe COVID-19. Conjunctival swab remains the gold standard of tear collection for RT-PCR assay. A significantly higher possibility of viral transmission exists through tears in patients with moderate to severe COVID-19. SARS CoV-2 viral RNA was detected in tears of 24% of laboratory proven moderate to severe COVID 19 patients, in early course of the disease, posing risk of viral transmission through tears. [ABSTRACT FROM AUTHOR]

The aim of the present study is to check the relationship between virus detection on the conjunctival swabs by RT-PCR and the systemic and ocular clinical data, treatments, and to the modalities of administration of supplemental oxygen. The SARS-CoV-2 RNA reverse-transcriptase PCR assay of conjunctival brushing samples and the corneal/conjunctival clinical findings were evaluated in 18 eyes of 9 consecutive patients admitted to the COVID-19 Sub-intensive Unit of Salerno Hospital University, Italy. Conjunctival swabs were positive for SARS-CoV-2 in 13 eyes of 7 patients; corneal epithelial defects were detected in 9 eyes. The seven patients with ocular involvement from SARS-CoV-2 had undergone treatment with a full-face mask or oxygen helmet in the last week, while the two subjects with negative conjunctival swabs had been treated with high flow nasal cannula. The positivity to the conjunctival test for SARS-CoV-2 was higher (72%) than that reported in the literature (10–15%) and related in all cases to the use of facial respiratory devices. These results suggest that exposure of unprotected eyes to aerosols containing high concentrations of SARS-CoV-2 could cause a keratoconjunctival viral infection. Further studies are needed to verify the causal link with the use of respiratory facial devices in patients suffering from COVID-19 pneumonia.

Conjunctival swabs (CS) are the most promising non-invasive samples for the diagnosis and the regular screening of Leishmania infantum infection in dogs although knowledge on their diagnostic performance is still inconclusive. This study evaluates CS real time-PCR (qPCR) analysis for the diagnosis of canine leishmaniosis (CanL) and its prognostic value in seropositive dogs from an endemic area. In October 2020 (T0), 26 dogs were enrolled, divided in two groups according to anti-L. infantum antibody titres (n = 13, group low titre (LT) and n = 13, group high titre (HT)), and followed-up in August 2021. At both timepoints, animals underwent clinical examination, complete blood count and biochemical analyses, and serological (indirect fluorescent antibody test) and molecular (CS and peripheral blood qPCR) testing. At T0, 10 out of 26 enrolled dogs were positive at CS qPCR, with the number of positive animals significantly higher in group HT than in LT. After 10 months, only 5 out of 21 dogs that completed the trial still tested CS qPCR positive, and none of them developed an active CanL based on clinical score and antibody titre. None of the dogs required any leishmanicidal and/or leishmaniostatic treatments. This prospective study showed unsatisfying diagnostic and prognostic performances of CS qPCR analysis in L. infantum seropositive asymptomatic dogs from an endemic area.

Purpose: The study objective was to identify severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in conjunctival swabs from patients with confirmed SARS-CoV-2 infection.Methods: Forty patients who tested positive by real-time reverse transcription (rRT)-polymerase chain reaction (PCR) of nasopharyngeal and oropharyngeal swabs were enrolled. Conjunctival swabbing was used to collect the tear and conjunctival secretions of patients.Results: Conjunctival swab rRT-PCR was positive for three patients and negative for 37 patients. Ten of the patients (25%) were diagnosed with conjunctivitis during the ophthalmic examination. Of these patients, one was found positive by conjunctival swab rRT-PCR, and nine were found negative. The difference between patients who tested positive or negative using conjunctival swab rRT-PCR was without statistical significance in terms of the presence of conjunctivitis (p = .720).Conclusion: The rate of positivity from conjunctival swab rRT-PCR was 7.5% in patients with confirmed SARS-CoV-2 infection. [ABSTRACT FROM AUTHOR]

Introduction. SARS-CoV-2 (severe acute respiratory syndrome-coronavirus-2) is a coronavirus that causes COVID-19 (coronavirus disease 2019) with mild to severe respiratory illness. It is a highly contagious disease transmitted through direct or indirect contact with infected people or contaminated surfaces, mainly through respiratory droplets, but other routes are being investigated. Objective. It is known that coronaviruses (CoVs) can cause a variety of ocular pathologies in animals, including conjunctivitis, anterior uveitis, retinitis, and optic neuritis, many of which are severe. However, there is no evidence of the SARS-CoV-2 presence in the eye tissue of asymptomatic patients, even if the symptomatic incidence is low. This systematic review presents updated literature on this issue. Abbreviated description of the state of knowledge. COVID-19 has now spread throughout the continents and poses a global threat to public health. The risk of rapidly overloading health care systems and causing substantial mortality worldwide is real. On 11 March 2020, the World Health Organization (WHO) announced coronavirus as a global pandemic. Several studies described a few cases with initial ocular symptoms followed by systemic symptoms of the disease. Summary. Although the frequency of transmission of SARS-CoV-2 infection through the eye is low, ocular symptoms are not uncommon in COVID-19. In some cases, eye symptoms may be the first signs of illness. This implies the need for hygienic recommendations and use of personal protective equipment (PPE) for medical staff and other services to minimize COVID-19 infection of both health-care workers and patients. A triage for ophthalmic outpatient clinic is mandatory. [ABSTRACT FROM AUTHOR]

The common approach of the diagnosis of Alzheimer’s Disease (AD) is made with an analysis of the cerebrospinal fluid or the study of retinal fundus and the plaques formation through optical corneal tomography (OCT), or more simply with a fundus camera. Tears analysis is widely discussed in literature as an essential method to describe molecular and biochemical alterations in different diseases. The aim of our study was the identification with immunocytochemistry of Amyloid Beta-42 in tears from patients with or without familiarity for Alzheimer Disease, in order to make the diagnosis earlier and more accessible compared to other invasive methods. Our study was performed on tears from three phenotypically healthy subjects: two of them were Caucasian with Alzheimer familiarity (48 and 55 years old) and the other one was Asian without Alzheimer familiarity (45 years old) and affected by an adenoviral keratoconjunctivitis at the moment of withdrawal. Tear samples were collected from eye fornix and were examinated by immunocytochemistry (ICC) assay using anti-Amyloid Beta X-42 antibody. Two out of three tears samples showed positive Amyloid Beta-42. Considering that our patients were phenotypically healthy, the identification of Amyloid Beta-42 by ICC could be a candidable method to make the diagnosis of the disease earlier and more accessible and available then other current and invasive methods and it could be a candidate for a screening method too.

Introduction: Purulent conjunctival discharge in hospitalized preterm infants may indicate conjunctivitis and warrant treatment. The purpose of this study was to examine the relationship between positive conjunctival swab (CS) culture and late-onset sepsis (LOS) in preterm infants., Methods: A retrospective cohort study was conducted to determine the relationship between positive CS culture growth results (CSP) obtained in preterm infants ≤34 weeks' gestation and the development of LOS within 120 h of obtaining CS compared with those who had negative CS culture results (CSN). Electronic medical records were reviewed from January 2015 until December 2019 for preterm infants presenting with purulent conjunctival discharge and underwent CS culture testing due to suspected conjunctivitis., Results: Of the 234 CS cultures obtained during the study period, 145 (61.9%) were CSP compared to 89 (38.1%) CSN cultures. Gram-negative organisms accounted for 70% of all CSP cultures, with the remaining 30% being Gram-positive. Patients with CSP were smaller, younger, had lower 1-minute APGAR scores, and required respiratory support more frequently than those with CSN. Infants with CSP received antibiotics for longer periods, both topically and systemically. Infants who developed LOS were more likely to require invasive ventilation (adjusted odds ratio, 33.5; 95% CI, 2.52-446.5, p  = 0.008). The incidence of LOS between the two groups was similar, with 6.2% observed in the CSP group compared to 3.4% in the CSN group ( p  = 0.543). Similarly, the rates of bacteremia were similar in both groups. Of the CSP patients who were presented with bacteremia, four out of seven (57%) exhibited bacteremia caused by the same organism found in their CS cultures. Similarly, within the entire cohort, respiratory cultures were performed on nine intubated patients within two weeks of obtaining CS cultures. Of these, in the CSP group, five out of six (83%) showed an organism identical to that found in the CS cultures., Conclusion: The study found a significant proportion of positive CS cultures in preterm infants, with distinct patient characteristics and treatment compared to negative cultures. While the incidence of LOS was not significantly different between the two groups, some CSP patients demonstrated bacteremia with the same CS organism, suggesting a possible connection between conjunctival or respiratory colonization and bacteremia., Competing Interests: No competing interests. All the authors are employed by Hamad Medical Corporation. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 Gad, Khalil, Halil, Chandra, Soliman, Rahman E'mar, Ibrahim, Al Khzzam, AlHendawi, Hamed, Bayoumi and Petkar.)

Abstract In Brazil, the visceral leishmaniasis (VL) is caused by Leishmania infantum , while the tegumentary leishmaniasis (TL) etiological agents are mainly Leishmania braziliensis and Leishmania amazonensis. The canine visceral leishmaniasis (CVL) diagnosis is an important step of the VL control program in Brazil, which involves the elimination of infected dogs, the main urban VL reservoirs. The current serology-based diagnostic tests have shown cross-reactivity between these three species, whereas molecular diagnosis allows high sensitivity and specie identification. In the present study, 349 dogs of the metropolitan region of Belo Horizonte (Minas Gerais state) were screened by conjunctival swab and the samples analyzed by ITS-1 nested PCR. Thirty dogs (8.5%) tested positive. The RFLP of amplicons using HaeIII demonstrated that 17/30 samples presented a banding pattern compatible with L. infantum , 4/30 matched with L. amazonenis , 1/30 with L. braziliensis and 8/30 showed a mixed infection pattern. The samples that were distinct of L. infantum or presented a mixed pattern were submitted to RFPL with HaeIII and RsaI enzymes that confirmed the mixed pattern. Such patterns were also confirmed by Sanger Sequencing. The results pointed eight dogs with mixed infections and the establishment of TL causing species in the Belo Horizonte dog population. These findings highlight the need for more comprehensive epidemiological studies, since the TL transmission profile might be changing. This study also shows the potential of the ITS1-nPCR associated with RFLP for the proper Leishmania diagnosis and typing in the dog population. [ABSTRACT FROM AUTHOR]

Purpose: To detect the presence of viral RNA of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in conjunctival swab specimens of coronavirus disease-19 (COVID-19) patients.Methods: Forty-five COVID-19 patients positive for real-time reverse transcription-polymerase chain reaction (RT-PCR) for SARS-CoV-2 in nasopharyngeal swab with or without ocular manifestations were included in the study. The conjunctival swab of each patient was collected by an ophthalmologist posted for COVID duty.Results: Out of 45 patients, 35 (77.77%) were males and the rest were females. The mean age was 31.26 ± 12.81 years. None of the patients had any ocular manifestations. One (2.23%) out of 45 patients was positive for RT-PCR SARS-CoV-2 in the conjunctival swab.Conclusion: This study shows that SARS-CoV-2 can be detected in conjunctival swabs of confirmed cases of COVID-19 patients. Though the positivity rate of detecting SARS-CoV-2 in conjunctival swabs is very less, care should be exercised during the ocular examination of patients of COVID-19. [ABSTRACT FROM AUTHOR]

Purpose The purpose of the study is to observe the characteristics of ocular manifestations in coronavirus disease 2019 (COVID-19) patients and to analyze the presence of severe acute respiratory syndrome coronavirus 2 ribonucleic acid (SARS-CoV-2 RNA) in the tears of patients with moderate-to-severe COVID-19. Material and methods We conducted this prospective cross-sectional study from February to June 2021 at the All India Institute of Medical Sciences, one of the tertiary eye care centers in Nagpur, India. The study included confirmed COVID-19 patients based on real-time reverse transcription-polymerase chain reaction (RT-PCR) nasopharyngeal swabs, whether or not the patients exhibited ocular symptoms. We recorded detailed information regarding the patients' history, including demographic profile, ocular symptoms, systemic symptoms, and radiologic findings. We collected ocular samples within 48 hours of collecting naso-oropharyngeal samples from the patients' eyes. We used conjunctival swabs to obtain tear samples, which we then placed in viral transport media (VTM) for cold chain transportation to the microbiology department. We performed RT-PCR on the tear samples to detect the presence of the SARS-CoV-2 virus. Result We included 40 patients in the study, with 26 (65%) classified as having moderate COVID-19, six (15%) classified as having severe COVID-19, and the remaining having mild COVID-19. Out of the 40 patients, five (12%) tested positive for SARS-CoV-2 in the tear sample using RT-PCR, seven (17%) exhibited ocular signs and symptoms, and only one tested positive for SARS-CoV-2 in their tears. The ocular manifestations observed in COVID-19 patients included dry eye, conjunctivitis (including conjunctival hyperemia and epiphora), and lid edema. Notably, we detected a positive COVID-19 tear sample in patients both with and without ocular symptoms. Conclusion Limited reports have focused on ocular involvement in patients with COVID-19. However, our study demonstrates the detection of SARS-CoV-2 in conjunctival swabs from confirmed COVID-19 patients, albeit with a lower positivity rate. Despite the low prevalence of the virus found in tears, there is a potential risk of transmission through ocular routes. It is noteworthy that we observed a COVID-19-positive tear sample in patients with and without ocular symptoms. Therefore, it is important to consider the possibility of ocular transmission even in the absence of ocular manifestations. Medical personnel should take careful precautions during ocular examinations of patients diagnosed with COVID-19 to minimize the risk of transmission., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Singh et al.)

The conjunctival sac of healthy human harbours a variety of microorganisms. When the eye is compromised, an occasional inadvertent spread happens to the adjacent tissue, resulting in bacterial ocular infections. Microbiological investigation of the conjunctival swab is one of the broadly used modality to study the aetiological agent of conjunctiva. However, most of the time such methods yield unsatisfactory results. Hence, the present study intends to identify the bacterial community in human conjunctiva of pre-operative subjects through 16S rRNA gene libraries. Out of 45 samples collected from preoperative patients undergoing cataract surgery, 36 libraries were constructed with bacterial nested-PCR-positive samples. The representative clones with unique restriction pattern were generated through Amplified Ribosomal DNA Restriction Analysis (ARDRA) which were sequenced for phylogenetic affiliation. A total of 211 representative clones were obtained which were distributed in phyla Actinobacteria , Firmicutes , α- Proteobacteria, β- Proteobacteria , γ- Proteobacteria, Bacteroidetes, and Deinococcus–Thermus. Findings revealed the presence of polybacterial community, especially in some cases even though no bacterium or a single bacterium alone was identified through cultivable method. Remarkably, we identified 17 species which have never been reported in any ocular infections. The sequencing data reported 6 unidentified bacteria suggesting the possibility of novel organisms in the sample. Since, polybacterial community has been identified consisting of both gram positive and gram negative bacteria, a broad spectrum antibiotic therapy is advisable to the patients who are undergoing cataract surgery. Consolidated effort would significantly improve a clear understanding of the nature of microbial community in the human conjunctiva which will promote administration of appropriate antibiotic regimen and also help in the development of oligonucleotide probes to screen the predominant pathogens for early predisposition. [ABSTRACT FROM AUTHOR]

Abstract: INTRODUCTION: Conjunctival swab PCR was evaluated as a tool to diagnose visceral leishmaniasis in dogs. METHODS: Conjunctival swab PCR was compared to indirect immunofluorescence antibody test and blood PCR. RESULTS: Indirect immunofluorescence was significantly correlated with conjunctival swab PCR (p < 0.05), but not with blood PCR (p > 0.05). In addition, conjunctival swab PCR was significantly associated with presence of clinical symptoms (p < 0.05), whereas blood PCR was associated with absence of clinical symptoms (p < 0.05). CONCLUSIONS: Results indicate that conjunctival swab PCR is useful in epidemiological surveys of canine visceral leishmaniasis.

Although some studies have investigated the potential role of cats as a reservoir for Leishmania , their role in the epidemiology of visceral leishmaniasis (VL) is still poorly understood. Molecular diagnostic techniques are an important tool in VL diagnosis, and PCR shows high sensitivity and specificity for Leishmania spp. detection. Quantitative real-time PCR (qPCR) is a method that permits quantitative analysis of a large number of samples, resulting in more sensitive, accurate, and reproducible measurements of specific DNA present in the sample. This study compared real-time PCR (qPCR) and conventional PCR (cPCR) for detection of Leishmania spp. in blood and conjunctival swab (CS) samples of healthy cats from a non-endemic area in the state of São Paulo, Brazil. Of all CS samples, 1.85% (2/108) were positive for Leishmania spp. by both cPCR as qPCR (kappa index = 1), indicating excellent agreement between the two methods. The DNA from the two CS-cPCR- and CS-qPCR-positive samples was further tested with a PCR test amplifying the Leishmania spp. discriminative rRNA internal transcribed spacer 1 (ITS 1), of which one sample generated a 300–350-bp DNA fragment whose size varies according to the Leishmania species. Following sequencing, the fragment showed 100% similarity to a GenBank L. infantum sequence obtained from a cat in Italy. In conclusion, the association of qPCR and CS proved to be effective for detection of Leishmania in cats. Conjunctival swab samples were shown to be a practical and better alternative to blood samples and may be useful in the diagnosis and studies of feline leishmaniasis. [ABSTRACT FROM AUTHOR]

Acanthamoeba is one of the most common free-living amoebas (FLA) that present in environment. In humans, Acanthamoeba can cause an infection of the eye termed Acanthamoeba keratitis, which mostly occurs in contact lens wearers. In the present study, we aimed to screen the presence of Acanthamoeba DNA in stray dogs using previously collected conjunctival swab samples in a hyper-endemic area for canine leishmaniasis. Totally, 184 dogs were included in the study and 27 of them (14.6%) were found positive for Acanthamoeba according to the 18s rRNA gene sequencing. Two different genotypes (T4 and T5) were identified and T5 was firstly reported in Turkey in the present study. Statistical analysis was performed and no correlation was found between Leishmania and Acanthamoeba positivity (P < 0.05). To best of our knowledge, this is the first study conducted to screen Acanthamoeba among stray dogs. Further studies are necessary to reveal the infection status and genotypes among dogs and its possible correlation with leishmaniasis. [ABSTRACT FROM AUTHOR]