Open Veterinary Journal, (2025), Vol. 15(12): 6351-6359

Research Article

10.5455/OVJ.2025.v15.i12.18

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Assessment of surface-associated bacterial contamination in Blatta orientalis from infested pig farms in Bulgaria

Betina Stefanova Boneva-Marutsova^*, Plamen Dimitrov Marutsov and Georgi Zhelev Georgiev

Department of Veterinary Microbiology, Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, Trakia University, Stara Zagora, Bulgaria

*Corresponding Author: Betina Stefanova Boneva-Marutsova. Department of Veterinary Microbiology, Infectious and Parasitic Diseases; Faculty of Veterinary Medicine, Trakia University, Stara Zagora, Bulgaria. Email: betina.boneva [at] trakia-uni.bg

Submitted: 08/05/2025 Revised: 30/10/2025 Accepted: 12/11/2025 Published: 31/12/2025

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Abstract

Background: Cockroaches are widely recognized as significant carriers of various infectious and parasitic diseases that affect both animals and humans. They are more than just common pests; they are among the most medically important insects due to their frequent presence in human environments. Understanding the role of cockroaches in disease transmission is crucial, and implementing effective pest control measures is essential.

Aim: This study focuses on identifying bacterial contaminants present on the surfaces of Oriental cockroaches (Blatta orientalis) collected from pig farms.

Methods: A total of 350 Oriental cockroaches collected from four field populations were examined in this study. Using the Vitek 2 Compact system, a modern platform for rapid and automated phenotypic identification, several bacterial species were isolated.

Results: A total of nine bacterial taxa were identified, with Escherichia coli being the predominant species (65.7% of pooled samples), followed by Staphylococcus spp., Streptococcus spp., Enterobacter spp., and Klebsiella pneumoniae. Other less frequent isolates included Pseudomonas spp., Enterococcus spp., Serratia marcescens, and Proteus mirabilis.

Conclusion: These results demonstrate that B. orientalis from pig farms harbors diverse bacterial contaminants of sanitary and veterinary concern, underlining the need for strict biosecurity and pest control measures to minimize the risk of disease transmission.

Keywords: Bacteria, Blatta orientalis, Cockroaches, Swine, Vitek.

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Introduction

Cockroaches are widely distributed insects of considerable economic, veterinary, and public health importance (Dent and Binks, 2020). Their extensive evolution has endowed them with remarkable adaptability, allowing survival in diverse environments, including extreme conditions, and enabling many species to thrive in human settlements, farms, and food industry facilities (Boneva-Marutsova, 2024). This synanthropic behavior raises critical concerns regarding their role in the transmission of infectious and parasitic (Alesho, 1997). Research has demonstrated that cockroaches are capable of carrying microorganisms and parasites on their surfaces, which positions them as significant vectors in the transmission of various infectious and parasitic diseases affecting both animals and humans (Vatev et al., 2006; Vahabi et al., 2007; Patel et al., 2022; Merad et al., 2023; Geng et al., 2025). Numerous studies identify them as primary carriers of food-borne pathogenic microorganisms (Gwenzi et al., 2021; Turner et al., 2023; Yulianti et al., 2023) and mechanical vectors (Luckyjane Molewa et al., 2022; Alhajeri et al., 2023; Davari et al., 2023; Abdullah et al., 2024). In light of the complex epidemic scenario related to the spread of African swine fever, an increasing number of researchers are raising concerns regarding the potential role of cockroaches and other insects in the dissemination of the disease (Fila and Woźniakowski, 2020; Yoon et al., 2020; Medrano, 2023).

Pathogens can remain on the surface of cockroaches or in their digestive systems for over a month after coming into contact with contaminated environments, food, or water. These pathogens may be released into the environment through their chitinous exoskeletons or feces (Moges et al., 2016; Liu et al., 2024). Given their role in disease transmission, assessing surface microbial contamination in cockroach populations cohabiting with humans and animals is crucial, and specialized cultivation and identification methods have been developed for this purpose (Fang et al., 2013; Moges et al., 2016; Solomon et al., 2016; Alikhani et al., 2017; Haile et al., 2018; Guzman and Vilcinskas, 2020; Chen et al., 2023; Davari et al., 2023). Among contemporary techniques enabling rapid and automated phenotypic identification of microorganisms through colorimetric analysis, the Vitek 2 Compact system (BioMérieux, France) is increasingly utilized in clinical practice (Alyas et al., 2021).

Our earlier studies on the distribution of cockroaches, conducted in different sites, established their presence in various livestock farms in Bulgaria. The highest prevalence of 100% was observed in industrial pig farms, where the degree of infestation was classified as high or very high. Particularly high levels of cockroach infestation have been observed in farrowing rooms, where they come into close contact with newborn piglets (Boneva-Marutsova, 2024), who are more susceptible to infectious diseases (Martínez-Boixaderas et al., 2022). The severity of the invasion raises concerns about its health implications regarding the spread of various infectious diseases. Currently, there is no information available about the bacterial carriage of the Oriental cockroach (Blatta orientalis) in intensive pig farms in Bulgaria, nor in many other countries in Eastern Europe. The aim of this study was to investigate bacterial surface carriage in field populations of cockroaches inhabiting pig farms in the Republic of Bulgaria to identify health and epidemic risks, using for the first time the VITEK 2 Compact system for their identification.

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Materials and Methods

A total of 350 Oriental cockroaches, collected from four industrial pig farms in Bulgaria heavily infested with high cockroach population densities, were examined in this study following the methodology of Solomon et al. (2016) and Davari et al. (2023). Sample collection was conducted between June and July 2023 during a single farm visit, employing live-capture methods. Cockroaches were trapped overnight and collected the next day with a specially adapted entomological bag. The highest population density was observed under the slatted floor of the farrowing house. Each farm yielded between 85 and 243 cockroaches, representing various developmental stages. Specimens were transported to the laboratory in boxes with tightly closing, locking lids, which were placed inside a larger container equipped with coolers to maintain suitable conditions during transit. During pool formation, all cockroaches that died during transport were removed. For each pool, cockroaches were randomly selected, excluding all immature stages. Due to an insufficient number of captured specimens, the number of pools was reduced to eight and seven for farms 2 and 4, respectively.

Following euthanasia by supercooling, groups of ten cockroaches were placed in sterile plastic containers containing 5 ml of sterile saline to create pooled samples. Surface contamination was extracted from each pool by vigorous vortexing at low speed (600–800 rpm) for 120 seconds, following the protocols of Alikhani et al. (2017) and Haile et al. (2018). From the supernatant, 1 ml was pipetted into two sterile Eppendorf tubes and transported under refrigeration to a specialized microbiological laboratory. Samples were inoculated onto blood agar (Blood Agar Base, HiMedia Laboratories, India) and MacConkey agar (HiMedia Laboratories, India), and incubated aerobically at 37°C for 24 hours (Dimri et al., 2020; Mariam, 2021). The isolates were identified using the automated densitometric VITEK 2 Compact System (BioMérieux Inc., France), which is increasingly used as a complementary analytical approach for identifying microbes in various samples from humans and animals, including cockroaches (Bizzini et al., 2011; Rettinger et al., 2012; Pérez-Sancho et al., 2015; Singhal et al., 2016; Manukumar and Umesha, 2017; Mehainaoui et al., 2021; Sánchez-Juanes et al., 2022). Upon examination of the Petri dishes, fresh pure monocultures (18–24 hours incubation) were prepared from colonies with distinct morphology. Following Gram staining, the appropriate identification card was selected: GP card for Gram-positive bacteria or GN card for Gram-negative bacteria. From each monoculture (2–3 well-developed colonies), standard suspensions of 0.50–0.63 McFarland were prepared using a DensiChek™ meter. Results were obtained with the VITEK 2 Compact System (BioMérieux, France) and automatically interpreted by the accompanying software. Quality control was ensured using reference strains.

Data analysis

All data were analyzed using IBM^® SPSS^® Statistics version 26.0. Categorical variables were compared among farms using the Chi-square test of independence. The strength of association between farm location and bacterial occurrence was evaluated using Cramér’s V coefficient, with thresholds interpreted as follows: 0.00–0.10 (negligible), 0.10–0.30 (weak), 0.30–0.50 (moderate), and >0.50 (strong association). Statistical significance was set at p < 0.05. Descriptive statistics (frequency, percentage) were used to summarize bacterial occurrence across farms.

Ethical approval

Not needed for this study.

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Results

Microbiological analysis aimed at evaluating the epidemiological potential of synanthropic cockroaches as mechanical vectors of pathogens of sanitary-hygienic and veterinary relevance revealed considerable bacterial presence on their external surfaces (Table 1).

Table 1. Surface bacterial contamination in Blatta orientalis collected from pig farms.

The quantitative distribution of GN and GP microorganisms isolated from the surface of synanthropic cockroaches collected from the surveyed pig farms is shown in Figures 1 and 2. The Cramér's V values, ranging from 0.092 to 0.482, indicate a weak to moderate association between bacterial species variability and farm location.

Fig. 1. Quantitative distribution of Gram-negative microorganisms isolated from Blatta orientalis across different pig farms.

Fig. 2. Quantitative distribution of Gram-positive microorganisms isolated from Blatta orientalis across different pig farms.

As shown in the summarized data in Figure 3, Escherichia coli was the predominant microbial isolate recovered from synanthropic cockroaches across all pig farms, followed by Staphylococcus spp., Streptococcus spp., and Enterobacter spp.

Fig. 3. Summary of the quantitative distribution of microorganisms isolated from Blatta orientalis across all pig farms.

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Discussion

Cockroaches are recognized as sanitary and hygienic pests of high epidemiological importance, acting as vectors for numerous pathogenic agents. When inhabiting contaminated environments or consuming infected food and water, various viruses, bacteria, fungi, and protozoa may persist on their exoskeletons or within their digestive tracts for extended periods and be subsequently released into the environment (Moges et al., 2016; Liu et al., 2024; Crespo et al., 2025).

Fang et al. (2013) and Chen et al. (2023) emphasize the critical need to assess microbial contamination in cockroach populations, particularly those cohabiting with humans and animals. Globally, most studies in this area have focused on German cockroaches and other synanthropic or exotic species. In Bulgaria, one of the few studies was conducted by Popova et al. (2010), also on an exotic cockroach species. However, there remains a substantial knowledge gap regarding the vectorial role of the black oriental cockroach, especially considering its frequent and severe infestations in pig farming facilities documented in our field observations.

In our investigation, significant surface bacterial contamination was detected in oriental cockroaches from pig farms (Table 1). The isolates included nine microbial taxa: Escherichia coli, Staphylococcus spp., Streptococcus spp., Enterobacter spp., Klebsiella pneumoniae, Pseudomonas spp., Enterococcus spp., Serratia marcescens, and Proteus mirabilis. These microorganisms are considered conditionally pathogenic, forming part of the normal or transient flora of the skin, mucous membranes, and intestines of mammals and birds, yet capable of causing disease under specific conditions in humans and animals. Our findings are consistent with previous studies, such as those by Zarchi and Vatani (2009), who identified 19 bacterial species from cockroach surfaces and digestive tracts, including E. coli, Group D Streptococcus, Bacillus spp., Klebsiella pneumoniae, and Proteus vulgaris. Similarly, Khodabandeh et al. (2020) reported a dominant presence of Klebsiella spp., Pseudomonas spp., Proteus spp., Citrobacter spp., Enterobacter spp., and Serratia spp. in German cockroaches.

A pronounced prevalence of Gram-negative bacteria, particularly from the Enterobacteriaceae family, was observed in our samples. This aligns with previous studies reporting similar bacterial profiles in German and American cockroach species and supports the potential role of cockroaches in spreading enteropathogens (Fotedar et al., 1991; Cloarec et al., 1992; Fotedar et al., 1992; Kobayashi et al., 1999; Pai et al., 2003; Magdy et al., 2023; Nirwan et al., 2024; Zainab Adesewa et al., 2024). Experimental studies by Zurek and Schal (2004) demonstrated that cockroaches shed large amounts of viable, virulent E. coli in their feces after initial exposure to contaminated material. Similarly, our study identified E. coli as the predominant contaminant on the surface of oriental cockroaches across all sampled farms. Pathogenic strains of E. coli are among the most frequently isolated pathogens in swine in veterinary practice (Fairbrother, 1999; Jacobson, 2022). Enterotoxigenic and verotoxigenic strains of E. coli are the main cause of colienteritis and colienterotoxicosis in newborn and weaned pigs, as well as edematous disease in postweaned pigs, often with high mortality (Bertschinger, 1999; Moon et al., 1999; Bassi et al., 2023). Waldvogel et al. (1999) reported the persistence of viable E. coli F18 strains in cockroach feces for up to 8 days’ post-exposure, with high bacterial loads (4.4 × 10⁵ CFU g⁻¹) comparable to those found in infected pigs (1.9 × 10⁶ CFU g⁻¹). This underscores the vector potential of cockroaches for these pathogens. Therefore, targeted cockroach density control is recommended as an integral part of disease prevention and control programs in pig farming (Waldvogel et al., 1999; Boneva et al., 2023).

Staphylococcus spp. and Streptococcus spp. were the next most frequently isolated genera in our study. Numerous investigations have highlighted the potential role of cockroaches as vectors of bacterial pathogens, particularly these two genera. Their frequent isolation from cockroach populations suggests a possible route for transmission to humans and animals. Studies by Fotedar et al. (1991) in India, Salehzadeh et al. (2007) in Iran, and Vazirianzadeh et al. (2009) have reported significant associations between cockroach infestations and the presence of pathogenic bacteria. Additionally, Schauer et al. (2012, 2014), Lampert et al. (2019), and Petrovskiy et al. (2024) identified high concentrations of staphylococci in various cockroach species, especially in the hindgut of Shelfordella lateralis. Of particular concern is S. aureus, a bacterium colonizing the skin and mucosal surfaces of approximately 30% of the human population, known for its ability to cause serious infections (Gorwitz et al., 2008). The increasing detection of antibiotic-resistant S. aureus strains in cockroaches, as documented by Menasria et al. (2014); Islam et al. (2016); Abdolmaleki et al. (2019); Akter et al. (2023); Yun et al. (2024), underscores their growing importance as a public health threat.

Our investigation revealed concerning levels of Klebsiella pneumoniae contamination across all surveyed pig farms. These findings are consistent with those of Cotton et al. (2000), who reported a high prevalence of Klebsiella in German cockroaches. Their work highlights the critical role of cockroaches as vectors in the transmission of nosocomial infections, emphasizing the urgent need for comprehensive pest control strategies in both agricultural and healthcare environments.

In one of the studied farms, Proteus mirabilis was detected on the surfaces of examined cockroaches. This bacterium has also been identified in the gastrointestinal tracts of German and American cockroaches (Robertson, 2007; Kundera et al., 2020), as well as in exotic species (Popova et al., 2010). Additionally, the presence of Pseudomonas aeruginosa was confirmed an opportunistic pathogen frequently isolated from various cockroach species, including those implicated in hospital infestations (Saitou et al., 2009). Pseudomonas aeruginosa is particularly notable for its high resistance to disinfectants and its prominent role in healthcare-associated infections.

The data unequivocally highlight cockroach monitoring and population control as crucial components of biosecurity programs, essential both for preserving animal health and for reducing the direct health risks to farm workers from potential exposure to cockroach-borne pathogens. The implementation of integrated pest management—encompassing physical, chemical, and organizational measures, alongside the maintenance of strict sanitary hygiene—is strongly recommended.

A potential limitation of this study is the lack of direct evidence demonstrating the transmission of pathogens between cockroaches and pigs on intensive farms, such as through molecular genetic analyses. Future research could address this gap by examining the impact of cockroach infestations on pig health across farms with varying levels of infestation (low, medium, high), focusing particularly on the incidence of neonatal diarrhea in farrowing facilities. The health risks associated with cockroaches, especially their role in maintaining and transmitting pathogens, become more pronounced during disease outbreaks. In such situations, control measures—like removing animals, cleaning feeders and waterers, and performing intensive disinfection with strong-smelling agents—often lead to the mass migration of cockroaches to nearby, unaffected facilities (Koehler et al., 2022). Therefore, implementing effective cockroach control should be an integral part of herd health management programs on pig farms. Conducting future in-depth studies would provide a more comprehensive understanding of the epidemiological significance of cockroaches and help inform targeted biosecurity and pest management strategies.

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Conclusion

The findings of this study highlight significant sanitary and epidemiological risks associated with cockroach infestations in livestock production and food processing environments. In particular, the detection of Blatta orientalis in swine facilities was strongly associated with high levels of bacterial surface contamination. Escherichia coli was identified as the predominant microbial isolate, followed by Staphylococcus spp., Streptococcus spp., Enterobacter spp., Klebsiella pneumoniae, Pseudomonas spp., Enterococcus spp., Serratia marcescens, and Proteus mirabilis, reflecting a diverse and complex microbial profile. These results underscore the necessity of implementing strict hygiene and biosecurity measures to mitigate the risk of pathogen transmission in such settings.

Conflicts of interest

The authors declare no conflicts of interest.

Funding

This research received no external funding.

Authors' contributions

Conceptualization, B.B-M., P.M. and G.Z.; methodology, B.B-M., P.M. and G.Z.; resources, B.B-M., P.M. and G.Z; writing—original draft preparation, B.B-M., P.M. and G.Z.; writing—review and editing, B.B-M., P.M. and G.Z.; visualization, B.B-M., P.M. and G.Z. All authors have read and agreed to the published version of the manuscript.

Data availability

The original contributions presented in this manuscript are included in the article. Further inquiries can be directed to the corresponding author.

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