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Volume 13
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10.3390/microorganisms13071487
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Article

Identification of Parasitic Infections by Analyzing Honeybees, Honey, and Pollen Using Droplet Digital RT-PCR

by
Luigi Jacopo D’Auria
1,†,
Andrea Mancusi
2,*,†,
Yolande Thérèse Rose Proroga
2,
Irene Dini
3,*,
Tiziana Cardellicchio
2,
Orlandina Di Maro
2,
Sabato De Vita
1,
Marica Egidio
4,
Raffaele Marrone
4 and
Giuseppe Rofrano
1
1
Centro di Referenza Nazionale per L’analisi e lo Studio delle Correlazioni tra Ambiente, Animali e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, 80055 Portici, Italy
2
Department of Food Security Coordination, Istituto Zooprofilattico Sperimentale del Mezzogiorno, 80055 Portici, Italy
3
Department of Pharmacy, University of Naples Federico II, 80138 Naples, Italy
4
Department of Veterinary Medicine and Animal Production, University of Naples Federico II, 80138 Naples, Italy
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Microorganisms 2025, 13(7), 1487; https://doi.org/10.3390/microorganisms13071487
Submission received: 26 May 2025 / Revised: 24 June 2025 / Accepted: 25 June 2025 / Published: 26 June 2025
(This article belongs to the Special Issue Advances in Genomics and Ecology of Environmental Microorganisms)
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Abstract

Toxoplasma gondii, Giardia intestinalis, and Cryptosporidium spp. are common pathogens that contaminate water and food. They can pose serious health risks, especially to vulnerable groups like immunocompromised individuals, pregnant women, young children, and aging people. An all-encompassing approach to minimizing transmission involves identifying effective techniques for detecting, treating, and preventing protozoan parasites. This study confirmed the effectiveness of a Droplet Digital Reverse Transcription Polymerase Chain Reaction (dd RT-PCR) method for quickly and accurately identifying Toxoplasma gondii, Giardia intestinalis, and Cryptosporidium species in honeybees, honey, and pollen by using ISO 17468 and ISO 16140 standard guidelines. The study evaluated honeybee (n = 16), honey (n = 12), and pollen (n = 8) samples collected from various apiaries in Southern Italy between June and September 2023. The results showed that honeybees, honey, and pollen can be considered bioindicators of infections by T. gondii, G. intestinalis, and Cryptosporidium spp. Furthermore, pollen, along with honey to a lesser degree, can serve as significant indicators for evaluating food safety. Therefore, it is essential to monitor their quality and purity due to environmental influences.
Keywords: bioindicators; droplet digital RT-PCR; hive products; honey; honeybees; pollen; protozoan parasites bioindicators; droplet digital RT-PCR; hive products; honey; honeybees; pollen; protozoan parasites

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MDPI and ACS Style

D’Auria, L.J.; Mancusi, A.; Proroga, Y.T.R.; Dini, I.; Cardellicchio, T.; Di Maro, O.; De Vita, S.; Egidio, M.; Marrone, R.; Rofrano, G. Identification of Parasitic Infections by Analyzing Honeybees, Honey, and Pollen Using Droplet Digital RT-PCR. Microorganisms 2025, 13, 1487. https://doi.org/10.3390/microorganisms13071487

AMA Style

D’Auria LJ, Mancusi A, Proroga YTR, Dini I, Cardellicchio T, Di Maro O, De Vita S, Egidio M, Marrone R, Rofrano G. Identification of Parasitic Infections by Analyzing Honeybees, Honey, and Pollen Using Droplet Digital RT-PCR. Microorganisms. 2025; 13(7):1487. https://doi.org/10.3390/microorganisms13071487

Chicago/Turabian Style

D’Auria, Luigi Jacopo, Andrea Mancusi, Yolande Thérèse Rose Proroga, Irene Dini, Tiziana Cardellicchio, Orlandina Di Maro, Sabato De Vita, Marica Egidio, Raffaele Marrone, and Giuseppe Rofrano. 2025. "Identification of Parasitic Infections by Analyzing Honeybees, Honey, and Pollen Using Droplet Digital RT-PCR" Microorganisms 13, no. 7: 1487. https://doi.org/10.3390/microorganisms13071487

APA Style

D’Auria, L. J., Mancusi, A., Proroga, Y. T. R., Dini, I., Cardellicchio, T., Di Maro, O., De Vita, S., Egidio, M., Marrone, R., & Rofrano, G. (2025). Identification of Parasitic Infections by Analyzing Honeybees, Honey, and Pollen Using Droplet Digital RT-PCR. Microorganisms, 13(7), 1487. https://doi.org/10.3390/microorganisms13071487

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