Prevalence and Antibiotic Resistance Pattern of Escherichia coli Isolated from Raw Dairy Milk

E. coli is one of the most important food borne pathogen, which could be transmitted by milk and milk products. To assess the role of dairy milk as the source of drug resistant E. coli, we examined 50 raw dairy milk samples (25-farm milk + 25-market milk) from some selected areas of Bangladesh by cultural, morphological, biochemical and antimicrobial sensitivity tests. In the preliminary observation, the mean total aerobic mesophilic count of market and farm raw milk samples were 8.98 and 8.68 log CFU/ml, while mean coliform count were 4.20 and 3.03 log CFU/ml respectively. Thirty-three E. coli isolates were recovered from collected samples (66% 33 of 50) and this pathogen was more prevalent in market milk (76%, 19 of 25) than farm milk (56%, 14 of 25). In addition, most of the isolated E. coli exhibited resistance against ampicillin and cefotaxime. This result shows that, the raw dairy milk and its products could be a source of human drug resistant E. coli

Molecular characterization and zoonotic risk assessment of Cryptosporidium spp. in children and calves in Bangladesh

Cryptosporidium is a gastro-intestinal protozoan parasite that has been found to infect both humans and livestock. This study investigated the parasite in 998 fecal samples from Bangladeshi children (n = 299) and calves (n = 699) to determine its prevalence, genetic variation, and zoonotic importance. The nested PCR and sequencing of the SSU rRNA gene in the samples showed a Cryptosporidium infection rate of 2.3% (7/299) in children and 15.7% (110/699) in calves. Statistical analysis revealed insignificant variations in Cryptosporidium infections among children across age, gender, and study area, while in calves, the infection rate significantly differed based on location and breed. Genotyping of seven human isolates of Cryptosporidium confirmed C. hominis (n = 5) and C. parvum (n = 2). After characterizing 110 Cryptosporidium isolates from calves, C. andersoni (n = 55), C. ryanae (n = 29), C. bovis (n = 14), C. parvum (n = 10), C. ubiquitum (n = 1), and C. occultus (n = 1) were identified. Cryptosporidium hominis and C. parvum-positive samples were further subjected to nested PCR and sequencing of the glycoprotein 60 (gp60) gene for subtyping. Four C. hominis subtypes (IaA19R3, IaA23R3, IbA9G3, and IdA15G1) and one C. parvum subtype (IIdA15G1) were observed. In conclusion, Cryptosporidium was prevalent in calves but less common in children in the study locations, and the presence of zoonotic Cryptosporidium species and subtypes in calves raises concerns regarding zoonotic transmission to humans

Predomination and new genotypes of Enterocytozoon bieneusi in captive nonhuman primates in zoos in China: high genetic diversity and zoonotic significance.

To appreciate the genetic diversity and zoonotic implications of Enterocytozoon bieneusi in nonhuman primates (NHPs) in zoos, we genotyped E. bieneusi in captive NHPs in seven zoos located at six major cities in China, using ribosomal internal transcribed spacer (ITS)-based PCR and sequence analyses. A total of 496 fecal specimens from 36 NHP species under nine families were analyzed and E. bieneusi was detected in 148 (29.8%) specimens of 25 NHP species from six families, including Cercopithecidae (28.7%), Cebidae (38.0%), Aotidae (75.0%), Lemuridae (26.0%), Hylobatidae (50.0%) and Hominidae (16.2%) (P = 0.0605). The infection rates were 29.0%, 15.2%, 18.2%, 37.3%, 29.2%, 37.7% and 44.8% in Shijiazhuang Zoo, Wuhan Zoo, Taiyuan Zoo, Changsha Wild Animal Zoo, Beijing Zoo, Shanghai Zoo and Shanghai Wild Animal Park, respectively (P = 0.0146). A total of 25 ITS genotypes were found: 14 known (D, O, EbpC, EbpA, Type IV, Henan-IV, BEB6, BEB4, Peru8, PigEBITS5, EbpD, CM1, CM4 and CS-1) and 11 new (CM8 to CM18). Genotype D was the most prevalent one (40/148), followed by CM4 (20/148), CM1 (15/148), O (13/148), CM16 (13/148), EbpC (11/148). Of them, genotypes D, EbpC, CM4 and O were widely distributed in NHPs (seen in 9 to 12 species) whereas genotypes CM1 and CM16 were restricted to one to three NHP species. In phylogenetic analysis, 20 genotypes (121/148, 81.8%), excluding genotypes BEB4, BEB6, CM9, CM4 and CM18, belonged to group 1 with zoonotic potential. New genotype CM9 clustered in group 2 with BEB4 and BEB6. The remaining two genotypes CM4 and CM18 formed new cluster (group 9) in between two other genotypic clusters found in primates. The findings of high diversity in E. bieneusi genotypes and their zoonotic potentiality concluded the importance of captive NHPs as reservoir hosts for human microsporidiosis

Genotype Analysis of Bacillus anthracis Strains Circulating in Bangladesh.

In Bangladesh, anthrax, caused by the bacterium Bacillus anthracis, is considered an endemic disease affecting ruminants with sporadic zoonotic occurrences in humans. Due to the lack of knowledge about risks from an incorrect removal of infected carcasses, the disease is not properly monitored, and because of the socio-economic conditions, the situation is under-reported and under-diagnosed. For sensitive species, anthrax represents a fatal outcome with sudden death and sometimes bleeding from natural orifices. The most common source of infection for ruminants is ingestion of spores during grazing in contaminated pastures or through grass and water contaminated with anthrax spores. Domestic cattle, sheep and goats can also become infected through contaminated bone meal (used as feed) originating from anthrax-infected carcasses. The present investigation was conducted to isolate B. anthracis organisms from 169 samples (73 soil, 1 tissue, 4 bone and 91 bone meal samples) collected from 12 different districts of Bangladesh. The sampling was carried out from 2012 to 2015. Twelve samples resulted positive for B. anthracis. Biomolecular analyses were conducted starting from the Canonical Single Nucleotide Polymorphism (CanSNP) to analyze the phylogenetic origin of strains. The analysis of genotype, obtained through the Multiple Locus Variable Number Tandem Repeat Analysis (MLVA) with the analysis of 15 Variable Number Tandem Repeats (VNTR), demonstrated four different genotypes: two of them were previously identified in the district of Sirajganj. The sub-genotyping, conducted with Single Nucleotide Repeats analysis, revealed the presence of eight subgenotypes. The data of the present study concluded that there was no observed correlation between imported cattle feed and anthrax occurrence in Bangladesh and that the remarkable genetic variations of B. anthracis were found in the soil of numerous outbreaks in this country

Neighbor-joining tree of <i>E</i>. <i>bieneusi</i> ITS genotypes.

<p>Phylogenetic relationship of <i>E</i>. <i>bieneusi</i> ITS nucleotide sequences of this study and other genotypes previously deposited in GenBank, as inferred by a neighbor-joining analysis (software Mega 5, <a href="http://www.megasoftware.net/" target="_blank">http://www.megasoftware.net/</a>) based on genetic distances calculated using the Kimura two-parameter model. The ITS tree was rooted with GenBank sequence DQ885585. Bootstrap values greater than 50% from 1,000 are shown on nodes. Each sequence from GenBank is identified by the accession number, host origin, and the genotype designation. The group terminology for the clusters is based on the works of Thellier and Breton [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0117991#pone.0117991.ref021" target="_blank">21</a>], Li et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0117991#pone.0117991.ref009" target="_blank">9</a>], and Karim et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0117991#pone.0117991.ref003" target="_blank">3</a>]. Two unique sequences of new genotype CM18 and known genotype CM4 in this study are designated as group 9 sequences. Known and new genotypes identified in this study are indicated by open and filled triangles, respectively.</p

Specific locations at which specimens were collected in this study.

<p>★locations of zoos.</p

Data on samples and sampling area.

<p>Data on samples and sampling area.</p

Map of Bangladesh indicating the origin of the samples.

<p><i>Image modified from “Map of Bangladesh” Author</i>: <i>Peter Fitzgerald</i>, <i>“World of Maps”- Public Domain (</i><a href="https://www.worldofmaps.net/asien/bangladesh/karte-regionen-bangladesch.htm" target="_blank"><i>https</i>:<i>//www</i>.<i>worldofmaps</i>.<i>net/asien/bangladesh/karte-regionen-bangladesch</i>.<i>htm</i></a><i>)</i></p

MLVA 15- VNTRs results.

<p>MLVA 15- VNTRs results.</p

Map of Bangladesh indicating the genotype distribution of <i>B</i>. <i>anthracis</i>.

<p><i>Image modified from “Bangladesh Administrative-2011”</i>, <i>803502AI (G00535) 6–11</i>. <i>Maps at the CIA—Public Domain (</i><a href="https://www.cia.gov/library/publications/resources/cia-maps-publications/index.html" target="_blank"><i>https</i>:<i>//www</i>.<i>cia</i>.<i>gov/library/publications/resources/cia-maps-publications/index</i>.<i>html</i></a><i>)</i>.</p