Genetic Polymorphism Characteristics of Brucella canis Isolated in China

In China, brucellosis is an endemic disease typically caused by Brucella melitensis infection (biovars 1 and 3). Brucella canis infection in dogs has not traditionally recognized as a major problem. In recent years however, brucellosis resulting from Brucella canis infection has also been reported, suggesting that infections from this species may be increasing. Data concerning the epidemiology of brucellosis resulting from Brucella canis infection is limited. Therefore, the purpose of this study was to assess the diversity among Chinese Brucella canis strains for epidemiological purposes. First, we employed a 16-marker VNTR assay (Brucella MLVA-16) to assess the diversity and epidemiological relationship of 29 Brucella canis isolates from diverse locations throughout China with 38 isolates from other countries. MLVA-16 analysis separated the 67 Brucella canis isolates into 57 genotypes that grouped into five clusters with genetic similarity coefficients ranging from 67.73 to 100%. Moreover, this analysis revealed a new genotype (2-3-9-11-3-1-5-1:118), which was present in two isolates recovered from Guangxi in 1986 and 1987. Second, multiplex PCR and sequencing analysis were used to determine whether the 29 Chinese Brucella canis isolates had the characteristic BMEI1435 gene deletion. Only two isolates had this deletion. Third, amplification of the omp25 gene revealed that 26 isolates from China had a T545C mutation. Collectively, this study reveals that considerable diversity exists among Brucella canis isolates in China and provides resources for studying the genetic variation and microevolution of Brucella.     

Viral and bacterial infections associated with camel (Camelus dromedarius) calf diarrhea in North Province,Saudi Arabia

Diarrhea and deaths in new-born camel calves were noticed by veterinary investigators and pastoralist in Saudi Arabia to be very high. Hence, it is thought to be necessary to investigate this problem from the virological and bacteriological point of view. The role of pathogenic bacteria and viruses in six different towns of North Province (Al-Assafia, Arar, Domat Aljandal, Hail, Skaka and Khoa) in Saudi Arabia was studied. Survey was conducted in diarrheic camel calves aged 12 months or younger. In our study calf diarrhea was reported in 184 out of 2308 camels examined clinically during one year, the prevalence of diarrhea was found to be 8.0% in calves ranging from one month to one year. In the present study group A rotavirus and Brucella abortus were detected in 14.7% and 8.98%, respectively, using ELISA technique. Escherichia coli was isolated from diarrheic calf camel (58.2%) 99/170 samples during dry and wet season. Salmonella spp. and Enterococcus spp. were detected in 12% and 8.8% of the specimens, respectively. In this study enterotoxogenic E. coli (ET E. coli) was isolated from 7% of diarrheic camel, which indicates the strong correlation between the camel calf diarrhea and the detection of enterotoxogenic E. coli. This study represented the first report for the detection of group A rotavirus and B. abortus antigen and antibodies in calf camels in Saudi Arabia. It is recommended that the disease should be controlled by vaccination in calf camels.     

Genetic variation and subspecific status of the grey wolf (Canis lupus) in Saudi Arabia

This work represents the most extensive genetic study of the grey wolf (Canis lupus Linnaeus, 1758) in Arabia and the first considering genetic data from multiple locations within Saudi Arabia. Previous suggestion of the occurrence of two subspecies of wolves in Arabia is not supported by this study. The genetic evidence suggests that the wolves of Saudi Arabia are genetically variable and more closely related to the Eurasian wolf Canis lupus group (dog included) than to the Indian wolf C. l. pallipes. The genetic diversity observed for C. lupus in Saudi Arabia indicates that the subspecific status C. l. arabs should be retained for the Arabian wolf. What remains unclear is the degree to which genetic introgression from domestic dogs has influenced the composition and integrity of C. lupus in Saudi Arabia.     

Amphibians distribution and habitats in the southwestern region of Saudi Arabia

This study surveyed 205 wetland sites in the Southwestern Saudi Arabia. We found seven species of amphibians (Anura), which belong to four families: Bufonidae; which includes four species, Sclerophrys tihamica; S. Arabica, Duttaphrynus dhufarensis, and Bufotes viridis; Hylidae, represented by only one species, the tree frog Hyla savignyi; Ranidae, which represented by one species, Pelophylax ridibundus and finally the Dicroglossidae, represented by one species, Euphlyctis ehrenbergii. The reasonable amount of rain received in southwest Arabia, and habitat diversity contribute to the relative abundance of amphibians in the region. Five types of wetland habitats were found in the study area. Valley streams, irrigated farms, seasonal ponds, dam reservoir and lagoons of treated sewage water. The current study revealed the wide spread of amphibians in southwestern Saudi Arabia than what had been reported earlier and confirmed the presence of the Bufotes viridis at three new sites in Asir Heights. It also showed the wide spread of S. arabica in all types of habitats in the southwestern region of Saudi Arabia. Habitat degradation was evident in the region, which might lead to species loss.     

Severity of Old World Cutaneous Leishmaniasis Is Influenced by Previous Exposure to Sandfly Bites in Saudi Arabia

BackgroundThe sandfly Phlebotomus papatasi is the vector of Leishmania major, the main causative agent of Old World cutaneous leishmaniasis (CL) in Saudi Arabia. Sandflies inject saliva while feeding and the salivary protein PpSP32 was previously shown to be a biomarker for bite exposure. Here we used recombinant PpSP32 to evaluate human exposure to Ph. papatasi bites, and study the association between antibody response to saliva and CL in endemic areas in Saudi Arabia.ConclusionsOur findings suggest a possible correlation between the type of immunity generated by the exposure to sandfly bites and disease outcome.     

Genetic diversity of Moringa peregrina species in Saudi Arabia with ITS sequences

The genus Moringa was the family of Moringaceae and Moringa oleifera and Moringa peregrina are the most famous species of Moringa. M. peregrina is widely grown in Saudi Arabia, Iran and India. Therefore, based on these reports, this study aimed to investigate the first systematic attempt to regulate the genetic diversity of the species M. peregrina in Saudi Arabian samples collected from several geographic locations using internal transcribed sequences. Genomic DNA was separated by CTAB extraction method and PCR was performed. Later on, DNA sequencing was performed for PCR products with ITS. In conclusion, the present study affords the first report on genetic stability of M. peregrina using ITS analysis in Saudi Arabia. Further studies are suggested in order to study in different regions.     

Tabanidae (Diptera) of Saudi Arabia

Samples were collected from southern, central and eastern regions of Saudi Arabia using Malaise traps and sweep nets. Nine species of Tabanidae were identified, two for the first time from Saudi Arabia, Hybomitra peculiaris (Szilády) and Atylotus pulchellus (Loew).Therefore, the total number of Tabanidae in Saudi Arabia is 31 species. Remarks of the species recorded in this study were given. A key to the genera of Tabanidae occurring in the Arabian Peninsula is also provided. Available literature for Saudi Arabian Tabanidae is summarized and provided. It is concluded that the tabanid fauna of Saudi Arabia is more similar to that of the Palaearctic region than to the Afrotropical region.     

Species-specific molecular signature of Commiphora species of Saudi Arabia inferred from internal transcribed spacer sequences of nuclear ribosomal DNA

The deciduous habit and tendency to produce flowers prior to developing leaves, and a predominantly dioecious system of breeding in the genus Commiphora leads to difficulties in its taxonomic identification at species level. The characteristics of easy amplification by universal primer, shorter length and higher discrimination power at the species level makes the internal transcribed spacer (ITS) sequence of nuclear ribosomal DNA (nrDNA) to a smart gene for generating species-specific phylogenetic inferences in most of the plants groups. The present study deals the ITS sequence of nrDNA based molecular genotyping of seven species of the genus Commiphora of Saudi Arabia. The molecular phylogenetic analysis of ITS sequences of nrDNA of Commiphora species distributed in Saudi Arabia reveals the the occurrence of C. madagascariens in Saudi Arabia.     

MLVA Genotyping of Brucella melitensis and Brucella abortus Isolates from Different Animal Species and Humans and Identification of Brucella suis Vaccine Strain S2 from Cattle in China

In China, brucellosis is an endemic disease and the main sources of brucellosis in animals and humans are infected sheep, cattle and swine. Brucella melitensis (biovars 1 and 3) is the predominant species, associated with sporadic cases and outbreak in humans. Isolates of B. abortus, primarily biovars 1 and 3, and B. suis biovars 1 and 3 are also associated with sporadic human brucellosis. In this study, the genetic profiles of B. melitensis and B. abortus isolates from humans and animals were analyzed and compared by multi-locus variable-number tandem-repeat analysis (MLVA). Among the B. melitensis isolates, the majority (74/82) belonged to MLVA8 genotype 42, clustering in the ‘East Mediterranean’ group. Two B. melitensis biovar 1 genotype 47 isolates, belonging to the ‘Americas’ group, were recovered; both were from the Himalayan blue sheep (Pseudois nayaur, a wild animal). The majority of B. abortus isolates (51/70) were biovar 3, genotype 36. Ten B. suis biovar 1 field isolates, including seven outbreak isolates recovered from a cattle farm in Inner Mongolia, were genetically indistinguishable from the vaccine strain S2, based on MLVA cluster analysis. MLVA analysis provided important information for epidemiological trace-back. To the best of our knowledge, this is the first report to associate Brucella cross-infection with the vaccine strain S2 based on molecular comparison of recovered isolates to the vaccine strain. MLVA typing could be an essential assay to improve brucellosis surveillance and control programs.     

First Record of the Myrmicine Ant Genus Meranoplus Smith, 1853 (Hymenoptera: Formicidae) from the Arabian Peninsula with Description of a New Species and Notes on the Zoogeography of Southwestern Kingdom Saudi Arabia

The ant genus Meranoplus is reported for the first time from the Arabian Peninsula (Kingdom of Saudi Arabia) by the new species M. pulcher sp. n., based on the worker caste. Specimens were collected from Al Sarawat and Asir Mountains of southwestern Kingdom of Saudi Arabia using pitfall traps. Meranoplus pulcher sp. n. is included in the Afrotropical M. magretii-group, with greatest similarity to M. magrettii André from Sudan. A key to the Afrotropical species of the M. magretii-group is presented. A brief review of the ant taxa with Afrotropical affinities in southwestern region of the Kingdom of Saudi Arabia is given.     

Modelling potential habitat suitability for critically endangered Arabian leopards (Panthera pardus nimr) across their historical range in Saudi Arabia

Camera trapping can detect and monitor rare species in landscapes spanning thousands of square kilometres but placement of cameras in areas where the animals most likely occur will increase detection success. This vital information is lacking for the critically endangered Arabian leopard (Panthera pardus nimr) that has undergone a 90% decline across its range in Saudi Arabia. We aimed to identify suitable Arabian leopard habitat and potential population capacity in Saudi Arabia using data from leopards living in ecologically analogous habitat in South Africa and Oman. We developed a resource selection function (RSF) from 14 leopards’ GPS data in the Cederberg, South Africa, and validated the model using three leopards in the Little Karoo, and two Arabian leopards in Oman. We then projected the model to the historical range of Arabian leopards in Saudi Arabia to estimate likely leopard locations and potential population sizes based on home range metrics. The RSF successfully discriminated between used and available locations (specificity = 96.7%) and had high predictive ability (Rho > 0.9). Leopards selectively used areas away from human settlements and roads, with high enhanced vegetation index, and intermediate slopes and elevations. Saudi Arabia could theoretically host 4 distinct populations totalling 162–362 Arabian leopard females, depending on home range size. Camera traps deployed in the south-western mountains of Saudi Arabia may be most likely to detect remnant populations of Arabian leopards. Further research is needed into the local abundance of prey species and human activity to ensure the persistence of suitable leopard ranges and inform conservation actions.     

Human Brucellosis in Maghreb: Existence of a Lineage Related to Socio-Historical Connections with Europe

Despite control/eradication programs, brucellosis, major worldwide zoonosis due to the Brucella genus, is endemic in Northern Africa and remains a major public health problem in the Maghreb region (Algeria/Morocco/Tunisia). Brucella melitensis biovar 3 is mostly involved in human infections and infects mainly small ruminants. Human and animal brucellosis occurrence in the Maghreb seems still underestimated and its epidemiological situation remains hazy. This study summarizes official data, regarding Brucella melitensis infections in Algeria, from 1989 to 2012, with the purpose to provide appropriate insights concerning the epidemiological situation of human and small ruminant brucellosis in Maghreb. Algeria and Europe are closely linked for historical and economical reasons. These historical connections raise the question of their possible impact on the genetic variability of Brucella strains circulating in the Maghreb. Other purpose of this study was to assess the genetic diversity among Maghreb B. melitensis biovar 3 strains, and to investigate their possible epidemiological relationship with European strains, especially with French strains. A total of 90 B. melitensis biovar 3 Maghreb strains isolated over a 25 year-period (1989–2014), mainly from humans, were analysed by MLVA-16. The obtained results were compared with genotypes of European B. melitensis biovar 3 strains. Molecular assays showed that Algerian strains were mainly distributed into two distinct clusters, one Algerian cluster related to European sub-cluster. These results led to suggest the existence of a lineage resulting from socio-historical connections between Algeria and Europe that might have evolved distinctly from the Maghreb autochthonous group. This study provides insights regarding the epidemiological situation of human brucellosis in the Maghreb and is the first molecular investigation regarding B. melitensis biovar 3 strains circulating in the Maghreb.     

Modeling,molecular docking,probing catalytic binding mode of acetyl-CoA malate synthase G in Brucella melitensis 16M

There are enormous evidences and previous reports standpoint that the enzyme of glyoxylate pathway malate synthase G (MSG) is a potential virulence factor in several pathogenic organisms, including Brucella melitensis 16M. Where the lack of crystal structures for best candidate proteins like MSG of B. melitensis 16M creates big lacuna to understand the molecular pathogenesis of brucellosis. In the present study, we have constructed a 3-D structure of MSG of Brucella melitensis 16M in MODELLER with the help of crystal structure of Mycobacterium tuberculosis malate synthase (PDB ID: 2GQ3) as template. The stereo chemical quality of the restrained model was evaluated by SAVES server; remarkably we identified the catalytic functional core domain located at 4^th cleft with conserved catalytic amino acids, start at ILE 59 to VAL 586 manifest the function of the protein. Furthermore, virtual screening and docking results reveals that best leadmolecules binds at the core domain pocket of MSG catalytic residues and these ligand leads could be the best prospective inhibitors to treat brucellosis.     

Survey of the reptilian fauna of the Kingdom of Saudi Arabia. V. The lizard fauna of Turaif region

Turaif area located in the Northern border region of Saudi Arabia is one of the most important regions of the Kingdom. This work was proposed to throw light on the diversity of lizard fauna investigated through the collection and subsequent identification of specimens from different localities of Turaif region of Kingdom of Saudi Arabia. Sixteen species of lizards belonging to 5 families (Agamidae, Gekkonidae, Lacertidae, Scincidae and Varanidae) were recorded. Lacertidae was the most common family. Three species of lizards namely Acanthodactylus orientalis, Acanthodactylus scutellatus and Acanthodactylus grandis were reported for the first time in the Turaif region of Saudi Arabia. The geographical distribution of the collected species within this province was mapped.     

Climate change and animals in Saudi Arabia

Global warming is occurring at an alarming rate and predictions are that air temperature (T_a) will continue to increase during this century. Increases in T_a as a result of unabated production of greenhouse gases in our atmosphere pose a threat to the distribution and abundance of wildlife populations worldwide. Although all the animals worldwide will likely be affected by global warming, diurnal animals in the deserts will be particularly threatened in the future because T_as are already high, and animals have limited access to water. It is expected that Saudi Arabia will experience a 3–5 °C in T_a over the next century. For predicting the consequences of global warming for animals, it is important to understand how individual species will respond to higher air temperatures. We think that populations will not have sufficient time to make evolutionary adjustments to higher T_a, and therefore they will be forced to alter their distribution patterns, or make phenotypic adjustments in their ability to cope with high T_a. This report examines how increases in T_a might affect body temperature (T_b) in the animals of arid regions. We chose three taxonomic groups, mammals, birds, and reptiles (Arabian oryx, Arabian spiny-tailed lizard, vultures, and hoopoe larks) from Saudi Arabia, an area in which T_a often reaches 45 °C during midday in summer. When T_a exceeds T_b, animals must resort to behavioral and physiological methods to control their T_b; failure to do so results in death. The observations of this study show that in many cases T_b is already close to the upper lethal limit of around 47° C in these species and therefore allowing their T_b to increase as T_a increases are not an option. We conclude that global warming will have a detrimental impact on a wide range of desert animals, but in reality we know little about the ability of most animals to cope with change in T_a. The data presented should serve as base-line information on T_b of animals in the Kingdom for future scientists in Saudi Arabia as they explore the impact of global warming on animal species.     

Identification, cloning and initial characterisation of FeuPQ in Brucella suis: a new sub-family of two-component regulatory systems

To cause disease, Brucella species have to adapt to a range of different environments. Environmental sensing and adaptive responses in bacteria often involve the concerted action of a two-component regulatory system, consisting of sensor and response regulator components. Amplification and sequence analysis of response regulators from Brucella species identified a response regulator sequence with 96% similarity to Rhizobium leguminosarum FeuP. In R. leguminosarum, the FeuPQ two-component system is involved in the regulation of iron uptake. A Brucella suis feuP isogenic mutant was constructed but was not attenuated in the murine brucellosis model. The survival and multiplication of the mutant in macrophages was also unaffected. The FeuPQ regulon represents a newly characterised sub-family of response regulators.     

Complete Genome Sequence of Brucella canis BCB018, a Strain Isolated from a Human Patient

Brucella canis is considered a rare cause of human brucellosis because of difficulties in presumptive diagnosis and underestimation of the incidence. Here, we report the draft genome sequence of a Brucella canis isolate, BCB018, isolated from a human patient, providing precious resources for comparative genomics analysis of Brucella field strains.     

Genotyping of Human Brucella melitensis Biovar 3 Isolated from Shanxi Province in China by MLVA16 and HOOF

Background

Brucellosis presents a significant economic burden for China because it causes reproductive failure in host species and chronic health problems in humans. These problems can involve multiple organs. Brucellosis is highly endemic in Shanxi Province China. Molecular typing would be very useful to epidemiological surveillance. The purpose of this study was to assess the diversity of Brucella melitensis strains for epidemiological surveillance. Historical monitoring data suggest that Brucella melitensis biovar 3 is the predominant strain associated with the epidemic of brucellosis in Shanxi Province.

Methods/Principal Findings

Multiple-locus variable-number repeat analysis (MLVA-16) and hypervariable octameric oligonucleotide fingerprinting (HOOF-print) were used to type a human-hosted Brucella melitensis population (81 strains). Sixty-two MLVA genotypes (discriminatory index: 0.99) were detected, and they had a genetic similarity coefficient ranging from 84.9% to 100%. Eighty strains of the population belonged to the eastern Mediterranean group with panel 1 genotypes 42 (79 strains) and 43 (1 strain). A new panel 1 genotype was found in this study. It was named 114 MLVAorsay genotype and it showed similarity to the two isolates from Guangdong in a previous study. Brucella melitensis is distributed throughout Shanxi Province, and like samples from Inner Mongolia, the eastern Mediterranean genotype 42 was the main epidemic strain (97%). The HOOF-printing showed a higher diversity than MLVA-16 with a genetic similarity coefficient ranging from 56.8% to 100%.

Conclusions

According to the MLVA-16 and HOOF-printing results, both methods could be used for the epidemiological surveillance of brucellosis. A new genotype was found in both Shanxi and Guangdong Provinces. In areas with brucellosis, the MLVA-16 scheme is very important for tracing cases back to their origins during outbreak investigations. It may facilitate the expansion and eradication of the disease.     

Complete Genome Sequence of Brucella melitensis 133, an Isolate of Biovar 1 of Sequence Type 32

Brucellosis is highly epidemic in China. Of the six classical species, Brucella melitensis and biovar 1 are the most represented species and biovar that cause human brucellosis in China. Here, we report the genome sequence of Brucella melitensis strain 133, a strain of biovar 1 of sequence type 32.     

Brucella melitensis Biovar 1 and Brucella abortus S19 Vaccine Strain Infections in Milkers Working at Cattle Farms in the Khartoum Area,Sudan

BackgroundHuman brucellosis is a preventable zoonoses that may become persistent, causing, if left untreated, severe localized disease. Occupational exposure to infected animals or animal products and consumption of fresh contaminated dairy are main risk factors.MethodsOne hundred farmworkers employed at two cattle farms one in Khartoum North and one in Omdurman were screened for the presence of specific antibodies and seropositive workers were invited to donate a blood sample for blood culture. Molecular typing was used to characterize Brucella isolates.ResultsTen percent of farmworkers tested seropositive and while Brucella melitensis biovar 1 was isolated from the blood of three individuals, an isolate identical to the B. abortus S19 vaccine strain was isolated from a fourth person. All four bacteremic individuals were employed as milkers and did not have obvious disease.ConclusionsThe isolation of the highly infectious pathogen B. melitensis from seropositive workers is consistent with the notion that the pathogen may persist in the blood without causing overt disease. While vaccination with strain S19 is essential for the control of bovine brucellosis the vaccine strain may be transmitted to the human population and protective measures remain important to prevent exposure also in view of the presence of B. melitensis. To create awareness for this potentially severe disease more information on the prevalence of the pathogen in different risk groups and in livestock in the Sudan is needed.