No abstract

Brucellosis is a major zoonotic infection in Kazakhstan. However, there is limited data on its incidence in humans and animals, and the genetic diversity of prevalent strains is virtually unstudied. Additionally, there is no detailed overview of Kazakhstan brucellosis control and eradication programs. Here, we analyzed brucellosis epidemiological data, and assessed the effectiveness of eradication strategies employed over the past 70 years to counteract this infection. We also conducted multiple loci variable-number tandem repeat analysis (MLVA) of Brucella abortus strains found in Kazakhstan. We analyzed official data on the incidence of animal brucellosis in Kazakhstan. The records span more than 70 years of anti-brucellosis campaigns, and contain a brief description of the applied control strategies, their effectiveness, and their impact on the incidence in humans. The MLVA-16 method was used to type 94 strains of B. abortus and serial passages of B. abortus 82, a strain used in vaccines. MLVA-8 and MLVA-11 analyses clustered strains into a total of four and seven genotypes, respectively; it is the first time that four of these genotypes have been described. MLVA-16 analysis divided strains into 28 distinct genotypes having genetic similarity coefficient that varies from 60 to100% and a Hunter & Gaston diversity index of 0.871. MST analysis reconstruction revealed clustering into "Kazakhstani-Chinese (Central Asian)", "European" and "American" lines. Detection of multiple genotypes in a single outbreak confirms that poorly controlled trade of livestock plays a crucial role in the spread of infection. Notably, the MLVA-16 profile of the B. abortus 82 strain was unique and did not change during 33 serial passages. MLVA genotyping may thus be useful for epidemiological monitoring of brucellosis, and for tracking the source(s) of infection. We suggest that countrywide application of MLVA genotyping would improve the control of brucellosis in Kazakhstan.

A c c e p t e d M a n u s c r i p t SummaryThis study was addressed to epidemiological surveillance of measles virus (MV) in the territory of Republic of Kazakhstan during 2015 -2016 years. In our study we have detected D8 genotype of MV occurred in outbreak season of measles disease. Totally 2341 cases were registered and 19 of them were identified by genotyping. As a result, 16 samples attributed to subgroup A of D8 genotype, and 3 import cases were represented by B3 and H1 genotype of MV. Vaccination coverage analysis showed that large group of infected people were not vaccinated and did not have a reliable report on vaccination status. This issue might increase the morbidity rate among the healthy population in outbreak season. To prevent the incidence caused by this problem we have successfully introduced necessary epidemiologic arrangements in controlling of measles.

Brucella abortus is the main causative agent of brucellosis in cattle, leading to severe economic consequences in agriculture and affecting public health. The zoonotic nature of the infection increases the need to control the spread and dynamics of outbreaks in animals with the incorporation of high resolution genotyping techniques. Based on such methods, B. abortus is currently divided into three clades, A, B, and C. The latter includes subclades C1 and C2. This study presents the results of whole-genome sequencing of 49 B. abortus strains isolated in Kazakhstan between 1947 and 2015 and of 36 B. abortus strains of various geographic origins isolated from 1940 to 2004. In silico Multiple Locus Sequence Typing (MLST) allowed to assign strains from Kazakhstan to subclades C1 and to a much lower extend C2. Whole-genome Single-Nucleotide Polymorphism (wgSNP) analysis of the 46 strains of subclade C1 with strains of worldwide origins showed clustering with strains from neighboring countries, mostly North Caucasia, Western Russia, but also Siberia, China, and Mongolia. One of the three Kazakhstan strains assigned to subclade C2 matched the B. abortus S19 vaccine strain used in cattle, the other two were genetically close to the 104 M vaccine strain. Bayesian phylodynamic analysis dated the introduction of B. abortus subclade C1 into Kazakhstan to the 19th and early 20th centuries. We discuss this observation in view of the history of population migrations from Russia to the Kazakhstan steppes.

I nfectious bovine keratoconjunctivitis (IBK), is a highly contagious and infectious disease of the bovine eye characterized by conjunctivitis and ulcerative keratitis that occurs worldwide (Holzhauer and Visser, 2004;Kneipp, 2021). Infectious keratoconjunctivitis also occurs in other domestic (Motha et al., 2003) and wild (Giacometti et al., 2002) species and is generally considered to be a multifactorial disease. The most commonly isolated pathogen from IBK is Moraxella bovis (M. bovis) (McConnel et al., 2007). The presence of M. bovoculi isolates in most eye samples taken from IBK-affected cattle suggests a role for M. bovoculi in pathogenesis (Angelos, 2010). The frequency of co-isolation of these two species may indicate that M. bovoculi serves as an opportunistic agent in IBK (Loy and Brodersen, 2014). In a recent study using the PCR method, M. bovoculi was detected in 75% of the samples of the total number of samples examined (Zheng et al., 2019). Other data on a positive correlation between the clinical signs of IBK and the presence of M. bovoculi also indirectly support that this organism is involved in the pathogenesis of IBK (Loy and Brodersen, 2014;Schnee et al., 2015).Pilin is known to be one of the key pathogenic factors in M. bovis, as this protein allows moraxella to "stick" to research Article Abstract | Moraxella bovoculi (M. bovoculi) is commonly isolated from clinical cases of infectious bovine keratoconjunctivitis (IBK), however, the exact role of M. bovoculi in this disease remains unclear. Experimental Moraxella bovis (M. bovis) and M. bovoculi vaccines to prevent IBK based on recombinant pathogenic factors have been evaluated as alternatives to whole cell vaccines. Pilin is one candidate antigen for M. bovoculi, and is a key pathogenic factor necessary for attachment of M. bovis to the surface of the eye epithelium. To evaluate pilA diversity amongst Kazakh isolates of M. bovoculi, the pilA gene sequence was determined for 35 isolates of M. bovoculi. Ten genotypes (gNA1-gNA10) were identified which were 96.1% to 99.8% identical. It was found that as a result of the replacement of cytosine with thymine at the 97th position of the gNA8 genotype, a stop codon resulted which, if expressed, would have resulted in a truncated PilA. Phylogenetic analyses of the deduced amino acid sequences of PilA from isolates from Kazakhstan, in comparison with previously defined PilA groups in M. bovoculi, established that 68% of the Kazakh isolates belonged to group A, 8.6% to groups C and F. It was found that despite these isolates originating from geographically close regions, some of the studied isolates had unique amino acid sequences that were assigned to new M. bovoculi PilA groups designated K (2 isolates), L (2 isolates), and M (1 isolate).