A flow cytometric method for enumeration and speciation of coccidia affecting broiler chickens

Adams, Daniel; Kulkarni, Raveendra R.; Mohammed, Javid P.; Crespo, Rocio

doi:10.1016/j.vetpar.2021.109634

Cited by 7 publications

(5 citation statements)

References 14 publications

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“…To date, there are a few automated protocols for oocyst enumeration and speciation using flow cytometry [9] and digital image analysis [10]. However, we found that the protocols used for sample preparation for these applications are complex, and the oocyst counts may be overestimated.…”

Section: Discussionmentioning

confidence: 92%

“…Automatic and semi-automatic protocols, including molecular tests, cytometry, and digital image analysis, for the enumeration and identification of coccidia are available [7][8][9][10]. Most PCR reactions target the intergenic transcribed spacer region 1 (ITS1) of the ribosomal RNA (rRNA) gene operon [7], a multi-copy gene, which makes these protocols unsuitable for estimation of the relative abundance of species in mixed infections of Eimeria sp.…”

Section: Introductionmentioning

confidence: 99%

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Automated Image Analysis for Detection of Coccidia in Poultry

Kellogg,

Roberts,

Crespo

2024

Animals

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Coccidiosis, caused by the protozoan Eimeria sp., is one of the most common and costly diseases impacting the poultry industry. To establish effective control measures, it is essential to identify these protozoa. Typical methods for identifying and determining the severity of the protozoal infection include intestinal lesion scoring or enumeration of the protozoal oocysts in fecal samples. Standard analysis methods require highly skilled technicians or veterinarians to manually identify and manually enumerate these protozoal parasites. This process is labor intensive, time-consuming, and susceptible to human error. None of the current methods available, including molecular flow cytometry or even digital image analysis, can determine if an oocyst is sporulated or not. Oocysts are not infectious until they sporulate. The goal of this study was to design an automated model using Artificial Intelligence (AI) to expedite the process of enumeration, improve the efficiency and accuracy of the species identification, and determine the ability of the oocysts to infect. To this end, we trained and evaluated computer vision models based on the Mask RCNN neural network architecture. A model was trained to detect and differentiate three species and to determine sporulation for each (totaling six detection groups). This model achieved a mean relative percentage difference (RPD) of 5.64%, representing a slight overcount compared to manual counts, averaging across all groups. The mean RPD for each group individually fell within a range from −33.37% to 52.72%. These results demonstrate that these models were speedy and had high agreement with manual counts, with minimal processing of field-quality samples. These models also could differentiate the sporulation status of the oocysts, providing critical diagnostic information for potential field applications.

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Section: Discussionmentioning

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Automated Image Analysis for Detection of Coccidia in Poultry

Kellogg,

Roberts,

Crespo

2024

Animals

Self Cite

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“…The poultry industry provides a large amount of meat and egg products for human consumption and its production scale is expected to further increase in the next decade ( Mottet and Tempio, 2017 ; Blake et al, 2020 ). Chicken coccidiosis is a widespread and economically significant disease caused by protozoan parasite of the genus Eimeria ( Chapman et al, 2013 ; Mesa et al, 2021 ), costing the global broiler industry more than 14 billion dollars per year ( Adams et al, 2022 ). There are 7 recognized Eimeria species, including E. Tenella, E. Acervulina, E. Maxima, E. Brunetti, E. Mitis, E. Necatrix , and E. Praecox .…”

Section: Introductionmentioning

confidence: 99%

A reliable and low-cost deep learning model integrating convolutional neural network and transformer structure for fine-grained classification of chicken Eimeria species

He¹,

Chen²,

He³

et al. 2023

Poultry Science

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“…Diseases caused by protozoa are also a challenge in animal husbandry, such as broiler chickens, laying hens or swine, at every stage of life. Each year, there are more than 14 billion-USD losses in the broiler chicken industry alone [ 3 , 4 , 5 ]. Moreover, some protozoa can be transmitted from animals to humans.…”

Section: Introductionmentioning

confidence: 99%

In Vitro Evaluation of Antiprotozoal Properties, Cytotoxicity Effect and Anticancer Activity of New Essential-Oil Based Phytoncide Mixtures

et al. 2023

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Protozoa, in both humans and animals, are one of the leading causes of disease. International programmes introduced in many countries have helped reduce the incidence of disease. However, it has recently become increasingly difficult to achieve the goals set for the coming years. One of the main reasons for this, as with other pathogenic organisms, such as bacteria and fungi, is the increasing resistance to current methods of treating and preventing infection. Therefore, new therapies with high efficacy are needed. In the present study, the novel mixtures of essential oils (EOs), clove, garlic, Ceylon cinnamon, and rosemary with organic acids (acetic, propionic, lactic) and metal ions (Cu, Mn, Zn) were tested against five selected model protozoa (Euglena gracilis, Gregarina blattarum, Amoeba proteus, Paramecium caudatum, Pentatrichomonas hominis). The cytotoxicity and potential anticancer activity of the obtained combinations were tested on the human fibroblasts (NHDF) and human cancer cell lines (A549, MCF7, LoVo, HT29). All of the mixtures showed very good antiprotozoal properties. The most efficient were the combination of clove and rosemary essential oils, mixtures of acids, and Mn ions. The LD50 values were in the range of 0.001–0.006% and the LD100 values were 0.002–0.008%. All of the tested mixtures did not show cytotoxicity against normal cells, but did show growth inhibition against cancer cell lines. The most cytotoxic against cancer cells were combinations with cinnamon essential oil. Nevertheless, the proposed combinations containing essential oils, organic acids, and metal ions have high antiprotozoal activity, with low toxicity to healthy human cells.

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A flow cytometric method for enumeration and speciation of coccidia affecting broiler chickens

Cited by 7 publications

References 14 publications

Automated Image Analysis for Detection of Coccidia in Poultry

Automated Image Analysis for Detection of Coccidia in Poultry

A reliable and low-cost deep learning model integrating convolutional neural network and transformer structure for fine-grained classification of chicken Eimeria species

In Vitro Evaluation of Antiprotozoal Properties, Cytotoxicity Effect and Anticancer Activity of New Essential-Oil Based Phytoncide Mixtures

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