Abstract
We report the incidence of bacteraemia associated withIgnatzschineria spp. for the first time from animal clinical case
presumably, as a post complication of maggot wound in a White Yorkshire
pig. We described a clinical history of a febrile adult white Yorkshire
pig and the isolation of Ignatzschineria spp. from blood sample.
The isolate was characterized phenotypically and further identified by
16S rRNA sequence analysis. Its occurrence may be misdiagnosed in
veterinary hospitals especially in low-resource settings, often leading
to the underreporting of such emerging infections, since the diagnostic
facilities are still in very primitive phase in developing countries.
More information on speciation is needed with much about the
epidemiology and pathogenesis of this emerging pathogen in order to
explore its role in the lives of animals and humans. Novel pathogens
continue to emerge in human, domestic animal, wildlife and plant
populations, yet the population dynamics of this kind of biological
invasion remain poorly understood. This rapid communication may redirect
the scientific community working for animal and human health worldwide
to unveil such rare emerging infections.
Keywords: Bacteraemia, Maggot infestation, Ignatzschineria spp.,
Myiasis in animals, Open wound, Emerging pathogen.
Introduction
Myiasis is an infestation of animals and humans by various dipteran
larvae, it was defined by Zumpt (1965) as “the infestation of live
vertebrate animals with dipterous larvae, which at least for a certain
period, feed on host’s dead or living tissues, liquid body substances,
or ingested food”. Such invasions may result in mild to severe illness
or even death. Flies of several families are obligatory parasites on
domestic animals, the most important family causing wound myiasis isCalliphoridae and included genera of flies are Calliphora,
Lucilia, Chrysomyia and Cochliomyia (Nadrah et al., 2021) but
flies belonging to family Calliphoridae and Sarcophagidaemostly cause traumatic wound and subsequent myiasis (Roy and Dasgupta,
1982) which is one of the commonest and widespread clinical problems in
veterinary practice in India (Chhabra & Pathak, 2009). Exposed lesions
attract and stimulate females to oviposit (Francesconi & Lupi, 2012).
Because flies and their larvae are the hosts to several bacterial
species, there are possibilities of secondary bacterial infections of
the wound and also bacteraemia sets in if invaded to the bloodstream.
The bacteria which are most frequently associated with myiasis-induced
bacteraemia are Wohlfahrtiimonas spp. and Ignatzschineriaspp. (Ahmad et al., 2022; Lysaght et al., 2018). Both the genera are
well known inhabitants of the salivary glands of larvae of many fly
species (Barker et al., 2014). The presence of Ignatzschineriaspp. has been reported from diverse environmental samples and animal
excreta (Juteau et al., 2004; De Luna et al., 2009). Here we describe
the very first incidence of myiasis with subsequentIgnatzschineria spp. bacteraemia from animal clinical case,
presumably as a post complication of maggot wound in an adult male of
White Yorkshire pigs.
Materials and Methods
In September 2021, a febrile male pig aging about 2 years of breed White
Yorkshire was presented with a history of pyrexia, cachexia and partial
anorexia from village Nardaha of Chhattisgarh state, India (Latitude:
21.2918203ON Longitude:
81.4313321OE). On physical examination it was found
that pig was wounded at the base of pinna and as per the history the
animal was suffered with maggot infested wound and treated with a
subcutaneous dose of Ivermectin @ 200 mcg/kilogram of body weight. The
animal belonged to a herd of about 250 animals, inclusive of piglets and
adult animals which were kept under semi-intensive production system.
Peripheral blood smear was prepared from febrile pig, air dried and
blood samples were collected aseptically. The collected samples were
transported to the laboratory on ice and processed for isolation of the
pathogens.
Sheep Blood agar (SBA) and MacConkey agar (MLA) were used as primary
culture media for preliminary isolation of organisms according to
methods described by Quinn et al. (2013). Briefly, blood samples were
streaked on SBA and MLA plates and incubated at 37oC
for 24 hours. Single colony of was picked up from primary culture and
re-streaked on fresh SBA plate and incubated at 37oC
for 24 hours to obtain pure culture. The organism was characterized by
Gram’s reaction, biochemical reactions and further identified by 16S
rRNA sequence analysis. The isolate was subjected to antibiotic
sensitivity testing by determination of minimum inhibitory concentration
(MIC). The EZY MIC™ Strips were obtained from HiMedia Laboratories Pvt.
Ltd., Mumbai, India. The isolates were tested against most commonly used
antibiotics and the results were interpreted as per CLSI 2020 guidelines
with breakpoints for other
non-Enterobacteriaceae applied. The antibiotics used for testing were
amoxycillin clavulanic acid, oxytetracycline, tetracycline, doxycycline,
kanamycin, gentamicin, vancomycin, rifampicin, cotrimoxazole,
ciprofloxacin, ofloxacin, pefloxacin, levofloxacin, cefotaxime,
cefotaxime-clavulanic acid, ceftriaxone, streptomycin and colistin.
Results and Discussions
Bacteriological cultures revealed as dominant growth of non-haemolytic,
smooth, greyish- white colonies of about 1 mm diameter size on sheep
blood agar (Fig. 1) and non-lactose-fermenting colonies on MacConkey
agar at 37˚C for 24 hours of incubation. Phenotypically, the organism
was Gram-negative bacilli (Fig. 2), positive for oxidase, catalase,
nitrate reduction and phenyl alanine deamination while negative for
urease, H2S production and did not utilize citrate, lysine, and
ornithine. The isolate could not able to ferment adonitol, arabinose,
cellobiose, glucose, dulcitol, galactose, inositol, inulin, lactose,
mannitol, mannose, raffinose, rhamnose, salicin, sorbitol and sucrose
and trehalose. Antibiotic Sensitivity Testing showed resistance towards
tetracycline group of antibiotics while showed susceptibility to all
other tested antimicrobials. According to the test results the animals
were treated with Enrofloxacin @ 5mg/Kg body weight per day for 5 days,
the animal was afebrile from the 2nd day onward. The
16S rRNA gene was amplified by PCR, and the amplicon underwent Sanger
sequencing (GCC Biotech Pvt. Ltd. Kolkata, India). The nucleotide
sequence data were analysed by BLAST (https://blast.ncbi.nlm.nih.gov),
which yielded 98.5% sequence homology to both Ignatzschineriaspp., assembled the genome sequences and submitted to GenBank under
accession numbers OP021690 and OP021691.This sequence was analyzed
on the nucleotide BLAST (BLASTn) suite of tools available on the NCBI
database (https://blast.ncbi.nlm.nih.gov/Blast.cgi). Default
parameters were employed to search the highly similar sequences among
16S rRNA genes in the database.