Rickettsia conorii subsp. israelensis infection in a pediatric patient presenting skin rash and abdominal pain: a case report from Southeast Iran.

BACKGROUND: The healthcare system in Iran appears to overlook Mediterranean spotted fever (MSF) as an endemic disease, particularly in pediatric cases, indicating the need for greater attention and awareness. CASE PRESENTATION: A six-year-old patient with fever, abdominal pain, headache, skin rashes, diarrhea, vomiting, and black eschar (tache noire) from southeast Iran was identified as a rickettsiosis caused by Rickettsia conorii subsp. israelensis through clinical and laboratory assessments, including IFA and real-time PCR. The patient was successfully treated with doxycycline. CONCLUSIONS: Symptoms like rash, edema, eschar, and abdominal pain may indicate the possibility of MSF during the assessment of acute febrile illness, IFA and real-time PCR are the primary diagnostic methods for this disease.

Molecular examination for Coxiella burnetii and Brucella spp. infections in Iranian women experiencing spontaneous miscarriage.

BACKGROUND: Spontaneous miscarriage, a leading health concern globally, often occurs due to various factors, including infections. Among these, Coxiella burnetii and Brucella spp. may have adverse effects on pregnancy outcomes. While previous research has established a link between infections and spontaneous miscarriage, our study aimed specifically to investigate the presence of these two pathogens in abortion samples from women who experienced spontaneous miscarriages in Iran. Our study can add to the existing knowledge by focusing on Iran, a region with a high prevalence of C. burnetii and Brucella spp. As a result, it could provide a better understanding and unique insights into the relationship of these pathogens with spontaneous miscarriages in endemic regions. METHODS: From March 2021 to March 2022, a total of 728 abortion samples (including placenta and cotyledon) were collected from 409 women who had experienced spontaneous miscarriages in the provinces of Tehran, Fars, and West Azerbaijan in Iran. The specimens included 467 Formalin-Fixed Paraffin-Embedded (FFPE) and 261 fresh frozen samples. After DNA extraction from abortion samples, the quantitative real-time PCR (qPCR) assay targeted a specific fragment of the IS1111 and IS711 elements for molecular identification of C. burnetii and Brucella spp., respectively. Furthermore, the qPCR assay employing specific primers for different species was used to determine the species of Brucella. RESULTS: Among the studied women, 1 out of 409 (0.24%) samples tested positive for Brucella spp., specifically Brucella melitensis. There were no positive specimens for C. burnetii. CONCLUSIONS: Our study contributes to understanding the potential involvement of Brucella species in spontaneous infectious abortion within endemic regions. The identification of B. melitensis in this study highlights the need for further research in this area. However, while our results suggest a relatively low or zero identification of these pathogens in our sample population, this does not rule out the possibility of undetected infections. Therefore, it is critical to acknowledge the limitations of the molecular techniques used (qPCR), which may have potential limitations such as sensitivity and specificity. Moreover, because 64.15% of our samples were FFPE, the sensitivity of the qPCR test may be reduced. These raise concerns about the accuracy of the reported prevalence rates and the potential for false positives or negatives.

Diversity of Rickettsia species in collected ticks from Southeast Iran.

Rickettsia occurs worldwide and rickettsiosis is recognized as an emerging infection in several parts of the world. Ticks are reservoir hosts for pathogenic Rickettsia species in humans and domestic animals. Most pathogenic Rickettsia species belong to the spotted Fever Group (SFG). This study aimed to identify and diagnose tick fauna and investigate the prevalence of Rickettsia spp. in ticks collected from domestic animals and dogs in the rural regions of Kerman Province, Southeast Iran. In this study, tick species (fauna) were identified and 2100 ticks (350 pooled samples) from two genera and species including Rhipicephalus linnaei (1128) and Hyalomma deteritum (972) were tested to detect Rickettsia genus using Real-time PCR. The presence of the Rickettsia genus was observed in 24.9% (95%CI 20.28-29.52) of the pooled samples. Sequencing and phylogenetic analyses revealed the presence of Rickettsia aeschlimannii (48.98%), Rickettsia conorii israelensis (28.57%), Rickettsia sibirica (20.41%), and Rickettsia helvetica (2.04%) in the positive samples. The results showed a significant association between county variables and the following variables: tick spp. (p < 0.001), Rickettsia genus infection in ticks (p < 0.001) and Rickettsia spp. (p < 0.001). In addition, there was a significant association between tick species and host animals (dogs and domestic animals) (p < 0.001), Rickettsia spp infection in ticks (p < 0.001), and Rickettsia spp. (p < 0.001). This study indicates a high prevalence of Rickettsia spp. (SFG) in ticks of domestic animals and dogs in rural areas of Kerman Province. The health system should be informed of the possibility of rickettsiosis and the circulating species of Rickettsia in these areas.

Development, Strategies, and Challenges for Tularemia Vaccine.

Francisella tularensis is a facultative intracellular bacterial pathogen that affects both humans and animals. It was developed into a biological warfare weapon as a result. In this article, the current status of tularemia vaccine development is presented. A live-attenuated vaccine that was designed over 50 years ago using the less virulent F. tularensis subspecies holarctica is the only prophylactic currently available, but it has not been approved for use in humans or animals. Other promising live, killed, and subunit vaccine candidates have recently been developed and tested in animal models. This study will investigate some possible vaccines and the challenges they face during development.

Fatal Case of Mediterranean Spotted Fever Associated with Septic Shock, Iran.

A fatal case of Mediterranean spotted fever associated with septic shock was reported in a 61-year-old man living in a village in southeastern Iran. The patient had a history of tick bite a few days before symptom onset. Phylogenetic analysis confirmed infection by Rickettsia conorii subspecies israelensis.

Rickettsia conorii subsp. israelensis infection: a case report from southeast Iran.

BACKGROUND: Mediterranean spotted fever (MSF) is a zoonotic and vector-borne disease caused by Rickettsia conorii. We report a case (36 year-old-woman) of MSF caused by Rickettsia conorii from Iran. CASE PRESENTATION: In September 2019, the patient was admitted to the hospital in Kerman province with flu-like symptoms and maculopapular lesions. According to the laboratory results, thrombocytopenia, elevated liver enzymes, and cardiac enzymes were observed. Skin biopsy was examined for Crimean-Congo Hemorrhagic Fever (CCHF) and MSF using the Real-Time-PCR and ELISA method. Finally, the sample was positive for Rickettsia conorii subsp. israelensis and treated with doxycycline and completely recovered. CONCLUSIONS: This study showed that MSF could be present in Iran. Therefore, identifying endemic areas in Iran for this disease should be on the agenda.

Tropheryma whipplei in the stool samples of children with acute diarrhea: a study from Tehran, Iran.

BACKGROUND: Recently, Tropheryma whipplei has been suggested as one of the causative agents of diarrhea among children worldwide. Limited data is available on the prevalence of T. whipplei among children with diarrhea in most countries such as Iran. This study was conducted to evaluate the prevalence of T. whipplei in children with acute diarrhea in Iran. METHODS: In this study, the stool samples were collected from 130 children under 10 years old with acute diarrhea from children's hospitals in Tehran city. Genomic DNA was extracted from stool samples and was tested for the presence of DNA of T. whipplei using the SYBR Green Real-time PCR method. Positive T. whipplei samples were finally confirmed by PCR Product sequencing. RESULTS: The mean age of participants was 32.5 months, and 54.6% of children were female. Using the SYBR Green Real-time PCR, 9.23% (12/130) of samples were positive for T. whipplei, which were confirmed by sequencing. 66.67% of positive cases were males. The duration of diarrhea in infected children with T. whipplei (83.3%) was significantly longer (OR: 5.93, 95% CI 1.24-28.22) compared to children with negative results (45.8%). Other demographic factors and clinical signs had not a statistically significant relationship with T. whipplei infection. CONCLUSIONS: In this study, T. whipplei was detected in stool samples of children with acute diarrhea. The results indicated that T. whipplei could be associated with childhood diarrhea in Iran. The health care system and physicians should be aware of the presence of T. whipplei infection in Iran, especially in childhood diarrhea.

The epidemiological survey of Coxiella burnetii in small ruminants and their ticks in western Iran.

BACKGROUND: Q fever is one of the most important zoonotic diseases caused by Coxiella burnetii. Although Q fever is an endemic disease in Iran, epidemiological data on C. burnetii infection are not yet complete in reservoirs and vectors in some parts of Iran. This survey investigated C. burnetii infection in small ruminants (sheep and goat blood samples) and their ticks in western Iran (Kurdistan province) in 2020. The presence of C. burnetii DNA was identified in these samples by targeting the IS1111 gene using the quantitative PCR (qPCR) method. RESULTS: Out of 250 blood samples (232 sheep and 18 goats), C. burnetii was detected in two samples (0.8%) belonging to the sheep (0.9%). In addition, 34 of 244 collected ticks (13.9%) from infested animals (244) were positive for C. burnetii infection. The highest prevalence of infection was found in Dermacentor marginatus (18.3%) and Haemaphysalis concinna (12.5%). CONCLUSIONS: The present study showed that ticks could have a possible role in the epidemiology of Q fever in Iran.

Targeted gene inactivation in Salmonella Typhi by CRISPR/Cas9-assisted homologous recombination.

BACKGROUND: Targeted gene inactivation (TGI) is a widely used technique for the study of genes' functions. There are many different methods for TGI, however, most of them are so complicated and time-consuming. New promising genetic engineering tools are developing for this purpose. In the present study, for the first time we disrupted a virulence gene from Salmonella enterica serovar Typhi (S. Typhi), located in the bacterial chromosome using CRISPR/Cas9 system and homology directed repair (HDR). METHODS: For this aim, pCas9 plasmid containing Cas9 enzyme and required proteins for homology directed recombination was transferred to S. Typhi by electroporation. On the other hand, a specific guide RNA (gRNA) was designed using CRISPOR online tool. Synthetic gRNA was cloned into pTargetF plasmid. Also, a DNA fragment (HDR fragment) was designed to incorporate into the bacterial chromosome following the cleavage of the bacterial genome by Cas9 enzyme. pTargetF containing gRNA and HDR fragment were co-transferred to S. Typhi containing pcas9 plasmid. The transformed bacteria were screened for recombination using PCR, restriction digestion and sequencing. RESULTS: The results of PCR, restriction digestion and sequencing showed the successful recombination of S. Typhi, in which the gidA gene is disrupted. CONCLUSION: In the present study we aimed to develop a rapid and robust method for targeted gene inactivation in a bacterial species, S. Typhi. This procedure can be exploited for disruption of other Salmonella as well as other bacteria's genes.

Francisella tularensis human infections in a village of northwest Iran.

BACKGROUND: Recent seroepidemiological studies have suggested that tularemia could be an endemic bacterial zoonosis in Iran. METHODS: From January 2016 to June 2018, disease cases characterized by fever, cervical lymphadenopathy and ocular involvement were reported in Youzband Village of Kaleybar County, in the East Azerbaijan Province, northwestern Iran. Diagnostic tests included Francisella tularensis serology (including tube agglutination test and ELISA), PCR, and culture. RESULTS: Among 11 examined case-patients, the tularemia tube agglutination test was positive in ten and borderline in one. PCR detected the F. tularensis ISFtu2 elements and fopA gene in one rodent and a spring water sample from the same geographic area. CONCLUSIONS: Based on the clinical manifestations of the disease suggesting an oropharyngeal form of tularemia, serology results in case patients, and F. tularensis detection in the local fauna and aquatic environment, the water supply of the village was the likely source of the tularemia outbreak. Intervention such as dredging and chlorination of the main water storage tank of the village and training of villagers and health care workers in preventive measures and treatment of the illness helped control the infection.

Molecular Detection of Zoonotic Pathogens in the Blood and Tissues of Camels (Camelus dromedarius) in Central Desert of Iran.

Dromedary camels (Camelus dromedarius) play a major economic role in many countries in Africa and Asia. Although they are resistant to harsh environmental conditions, they are susceptible to a wide range of zoonotic agents. This study aimed to provide an overview on the prevalence of selected zoonotic pathogens in blood and tissues of camels in central Iran. Blood, liver, portal lymph node, and brain were collected from 100 apparently healthy camels at a slaughterhouse in Qom city to assess the presence of DNA of Brucella spp., Trypanosoma spp., Coxiellaburnetii, and Bartonella spp. PCR products were sequenced bidirectionally and phylogenetic analyses were performed. Eleven percent of camels tested positive for Brucellaabortus (3%) and Trypanosomaevansi (8%). Coxiellaburnetii and Bartonella spp. DNA was not detected. Our data demonstrate that camels from Iran contribute to the epidemiology of some zoonotic pathogens. Performing proper control strategies, such as vaccination of camels and humans in contact with them, test-and-slaughter policy, and education of the general population is necessary for minimizing the risk of zoonotic infection.

Serological evidence of leptospirosis in Iran; A systematic review and meta-analysis.

Leptospirosis is a neglected infectious zoonotic disease that affects domestic animals and wildlife as well as humans. Although leptospirosis is known as an endemic disease in Iran, there is no accurate information on the overall prevalence of this disease in humans and animals. The aim of this systematic review and meta-analysis was to estimate the seroprevalence of leptospirosis among human and domestic and wild animals in Iran. A systematic review of English and Persian articles (since 1998 to December 2017) was conducted using Google Scholar, Medline/PubMed, Science Direct, Scopus, Web of science and Iranian databases Iranmedex, Scientific Information Database (SID), Magiran, and IRANDOC. Search terms include leptospirosis, Leptospira, serology, seroprevalence, seroepidemiology, serological, Iran, cow, goat, sheep, camel, dog, cat, equine, donkey, horse, mule and rodent. In Eventually 66 articles were selected to analyze based on inclusion criteria. Seroprevalence of leptospirosis in human was 27.84% (95% CI: 13.22-22.47) and 19.71% (95% CI: 6.78-32.65%) based on ELISA and MAT, respectively. The pooled prevalence of leptospirosis in cow, sheep, goat and camel was 26.62% (95% CI: 18.76-34.48), 17.38% (95% CI: 13.32-21.43), 12.18% (95% CI: 9.96-14.41) and 22.68% (95% CI: 18.97-26.40), respectively. The prevalence of leptospirosis in horse, donkey, and mule was 19.99% (95% CI: 13.32-26.68), 40.59% (95% CI: 33.20-47.97) and 9.10% (95% CI: 2.90-15.30), respectively. The prevalence in dog and cat were estimated 14.63% (95% CI: 3.49-25.77) and 14.44% (95% CI: 3.25-25.65), respectively. The prevalence of seropositivity in rodents was estimated 20.96% (95% CI: 10.62-31.30). This study is a very comprehensive report on the status of leptospirosis in Iran. Based on our results, leptospirosis has considerable seroprevalence among human and animals in Iran. This high seroprevalence of leptospirosis showed should be given more attention for this disease in Iran and thus health measures must be taken to diagnosis, control and prevent it.

Epidemiological survey of tularemia in Ilam Province, west of Iran.

BACKGROUND: Francisella tularensis is the causative agent of tularemia in humans and a large number of animal species. Considering recent evidence of the circulation of this bacterium in different parts of Iran, especially in the western provinces, the aim of current study was to determine the tularemia seroprevalence in the human population living in Ilam Province. METHODS: In 2015, 360 serum samples were collected from five groups of people: ranchers (n = 112), farmers (n = 79), butchers and slaughterhouse workers (n = 61), Nature Conservation Officers (n = 34), and referents of medical diagnostic laboratories (n = 74). These samples were tested for the presence of anti- F. tularensis IgG antibodies using the ELISA method. RESULTS: According to the ELISA manufacturer cutoffs, we found that 10 (2.78%) and 9 (2.5%) sera, respectively, were positive or borderline for F. tularensis IgG antibodies. The highest tularemia seroprevalence was observed among farmers (7.59%). CONCLUSIONS: Our results strongly support the circulation of tularemia in Ilam Province. Because no human tularemia case has been reported so far in this province, we recommend specific education programs to increase knowledge of local health care professionals about this important zoonotic disease.

Molecular prevalence of Coxiella burnetii in milk in Iran: a systematic review and meta-analysis.

Q fever is a major zoonotic disease in the world. The aim of this meta-analysis was to estimate the prevalence of Coxiella burnetii in animal milk in Iran. We systematically reviewed the literature to identify eligible studies from January 2008 to June 2016 in English or Farsi (Persian) databases. We extracted the molecular prevalence of C. burnetii in milk from cows, goats, sheep, and camels in Iran. The total prevalence of C. burnetii in cow milk was 15.09% (95% CI 11.08-19.10) by PCR methods. The highest and lowest prevalence of Q fever agent were seen in the East Azerbaijan (25.55%) and Khorasan-Razavi (4.22%) provinces, respectively. The molecular prevalence of C. burnetii in goat milk was 7.80% (95% CI 3.54-12.07%). The provinces of Qom (0%) and Lorestan (44.71%) had the lowest and the highest frequency of C. burnetii infection in goat's milk, respectively. Total prevalence of C. burnetii in sheep milk was 3.79% (95% CI 0.72-6.87%). The highest frequency of C. burnetii in sheep milk was detected in the Khorasan-Razavi province (34.78%). The frequency of C. burnetii in camel milk was 1.43%. High infection of C. burnetii in milk is an important health problem in Iran, amplified by the traditional preparations of dairy products.

Epidemiology of tularemia in the countries of the WHO Eastern Mediterranean Region (EMRO): A systematic review and meta-analysis.

BACKGROUND: Francisella tularensis, the bacterium that causes tularemia, has been a persistent and widespread pathogen in various regions of the world for centuries. Francisella tularensis can affect humans and various domestic and wild animals. The current study aimed to determine the epidemiological status of tularemia in countries of the WHO Eastern Mediterranean Region (EMRO) through a systematic review and meta-analysis. METHODS: All included studies were identified through a systematic search of online databases, including Scopus, PubMed, Web of Science, and EMBASE, through July 26, 2022, using keywords and suitable combinations. We focused on cross-sectional studies investigating the prevalence of F. tularensis. The weighted pooled prevalence was calculated using a random-effects model. RESULTS: A total of 206 studies were identified, of which 20 were finally included in the analysis. The human seroprevalence of tularemia in WHO-EMRO countries was 6.2% (95% CI, 4.2 9.2). In the subgroup analysis, anti-F. tularensis antibodies were found in 6.92% and 5.5% of the high-risk individuals and Iran, respectively. The pooled prevalence of F. tularensis in environmental samples (water and soil) from the WHO-EMRO countries was 5.8% (9.4% by PCR and 0.5% by culture). In addition, 2.5% (95% CI, 0.2 0.22.7) of ticks in WHO-EMRO countries were positive for F. tularensis. The pooled prevalence of F. tularensis in rodents is 2.0% (1.1% by PCR and 3.7% by serology). In addition, 0.6% of domestic ruminants (0.4% by PCR and 2.4% by serology) were positive for F. tularensis in WHO-EMRO countries. CONCLUSION: According to the results of the present study, tularemia is an endemic but neglected disease in the WHO-EMRO region. However, most studies on tularemia are limited to a few countries in this region. Studies on tularemia in human populations, reservoirs, and vectors have been conducted in all countries in the WHO-EMRO region to obtain more detailed information about the epidemiology of tularemia in these regions.

Distribution of different Rickettsia species in countries of the WHO Eastern Mediterranean (WHO-EMRO) region: An overview.

SUBJECT: Rickettsia is a zoonotic bacterial pathogen transmitted by vectors and has extensive reservoirs in animal and human populations. Rickettsiosis is a public health problem all over the world. However, comprehensive information on the geographical distribution of different Rickettsia species, infection status of reservoirs, vectors, and human cases is lacking in most parts of the world. Therefore, this study aimed to investigate the geographical distribution of different Rickettsia species and their vectors in countries of the WHO-EMRO region. METHODS: In this review study, a search was conducted for reports and published studies on Rickettsia species from WHO-EMRO region countries in various databases from 1995 to 2022. Finally, the reported status of human cases, reservoirs, and vectors associated with each species in different countries was documented. RESULTS: Reports of infections related to the detection of Rickettsia species were only available for 15 out of 22 WHO-EMRO member countries. A total of twenty-four Rickettsia species, including R. sibrica, R. lusitaniae, R. africae, R. prowazekii, R. felis, R. typhi, R. rickettsii, R. aeschlimannii, R. conorii, R. massiliae, R. helvetica, R. monacensis, R. rhipicephali, R. bellii, R. asembonensis, R. hoogstraalii, R. andeanae, R. raoultii, R. asiatica, R. slovaca, R. australis, R. barbariae, Candidatus R. amblyommii, and Candidatus R. goldwasserii, were reported from WHO-EMRO member countries. Furthermore, human cases infected with six different Rickettsia species, including R. sibrica, R. prowazekii, R. felis, R. typhi, R. rickettsii, R. aeschlimannii, R. conorii, R. massiliae, and R. helvetica, were reported from these countries. CONCLUSION: The vast diversity of Rickettsia vectors has contributed to the ongoing discovery of new Rickettsia species. Therefore, further research on the reservoir hosts of Rickettsia infections in the understudied WHO-EMRO region is crucial. This research sheds light on Rickettsia disease's epidemiology and transmission dynamics in this region.

Molecular investigation of Coxiella burnetii in aborted fetus of small ruminants in southeast Iran.

The domestic animal, known as a main reservoir of Coxiella burnetii, is susceptible to the occurrence of coxiellosis, which can lead to abortions in domestic animals, causing significant economic damage and posing risks to human health. Therefore, the purpose of this study is to investigate C. burnetii as the causative agent of Q fever in abortion samples of small ruminants in southeastern Iran. This study was conducted between 2020 and 2021 in Zarand city, located in Kerman province (southeast Iran). In this study, 50 abomasum swab samples of aborted sheep and goat fetuses were collected and analyzed using molecular methods to identify C. burnetii. The results revealed that 26% (n: 13) of the collected abortion samples were infected with C. burnetii. Among the positive samples, two (50%) belonged to goat abortion samples while 11 (23.9%) belonged to sheep abortion samples. This study demonstrates that C. burnetii is one of the causes of abortion in small ruminants in southeastern Iran. It is recommended to pay more attention to C. burnetii in domestic animals due to its significant economic impact on livestock and its potential implication for human health in Iran.

Geographical distribution of Bartonella spp in the countries of the WHO Eastern Mediterranean Region (WHO-EMRO).

Bartonellosis is a vector-borne and zoonotic diseases in humans, especially in immunocompromised individuals. However, there is no complete data about the geographical distribution of different species of Bartonella, as well as the status of its reservoirs, vectors, and human cases in most parts of the world. In this study, published reports related to Bartonella species from WHO-EMRO region countries were searched in different databases until October 2023. The eighteens different species of Bartonella were reported in WHO-EMRO countries including Bartonella henselae, Bartonella quintana, Bartonella elizabethae, Bartonella bovis, Bartonella clarridgeiae, Bartonella vinsonii, Bartonella doshiae, Bartonella taylorii, Bartonella rochalimae, Bartonella tribocorum, Bartonella rattimassiliensis, candidatus Bartonella merieuxii, candidatus Bartonella dromedarii, Bartonella acomydis, Bartonella jaculi, Bartonella coopersplainsensis and Bartonella koehlerae. Also, only human cases of B. henselae and B. quintana infections were reported from WHO-EMRO countries. The infections of Bartonella are important in the WHO-EMRO region, but they have been neglected by clinicians and healthcare systems.

Molecular detection of Brucella species among aborted small ruminants in southeast Iran.

Brucellosis, caused by Brucella bacteria, is a common zoonotic infectious disease with various clinical manifestations in humans and animals. The disease is endemic in human and ruminant populations in Iran, with a particular prevalence in areas where humans have close interactions with livestock. Since domestic animals serve as the primary reservoir for brucellosis, this study aimed to identify the presence of Brucella spp. among aborted small ruminants in southeast Iran. Between 2021 and 2022, aborted fetuses of small ruminants (46 sheep and 4 goats) were collected from Zarand County in the Kerman province. Swab samples from the abomasum contents of these fetuses were obtained and subjected to DNA extraction. The samples were then tested for Brucella spp. detection using the polymerase chain reaction (PCR) method. Out of the 50 aborted fetuses examined, Brucella spp. was detected in 15 (30%) specimens, comprising 13 (28%) sheep and 2 (50%) goats. Species typing revealed the presence of Brucella ovis (6 sheep and 1 goat), Brucella melitensis (6 sheep), and Brucella abortus (1 sheep) among the positive specimens. This cross-sectional study highlights the high prevalence of various Brucella species in samples from small ruminant abortions in southeast Iran. Additionally, the identified Brucella species were not limited to their primary host livestock. These indicated potential cross-species transmission among small ruminants.