Spatial distribution, host specificity, and affinity of Branchiura for surface and macrohabitat types in fish species from the Brazilian Amazon.

This study investigated the spatial distribution, parasite interactions, and affinities of Branchiura parasites according to macrohabitat and surface type on fish from the Araguari River, Amapá State, in the Brazilian Amazon. This study presents novel information on the interactions of branchiuran species with host fish, and how the morphology of their attachment structures is associated with different host surface morphologies, with and without scales. A total of 558 fish specimens of 26 species were collected, of which 109 fish (10 species) were parasitized by at least one branchiuran parasite; these were selected for further study. The parasites were collected from the buccal cavity, gills, fins, and integument of the host fish. Parasites with suction discs (Argulus) were mainly associated to hosts with small scales and no scales, while parasites with hooks (Dolops) were mainly associated to hosts with large scales. Overall, parasites with suction discs were more specialized per site than those with hooks.

Lethal concentration of Cymbopogon citratus (Poaceae) essential oil for Dolops discoidalis and Argulus sp. (Crustacea: Argulidae).

This study investigated the acute toxicity (LC50-24 hr ) effects of the essential oil of Cymbopogon citratus for adult Argulus sp. and Dolops discoidalis, before and during oviposition. In vitro acute toxicity (LC50-24 hr ) was tested using 20, 40, 60, 80, 100, 120, 140 and 160 µg/L of C. citratus essential oil, and two control groups (one with cultivation tank water and one with cultivation tank water + alcohol) were used. Specimens of Argulus sp. and D. discoidalis submitted to acute toxicity were evaluated using histological procedures. The major chemical compounds of C. citratus essential oil were geranial (47.5%), neral (35.6%) and myrcene (6.7%). The LC50-24 hr for Argulus sp. adults was 67.97 µg/L, while for D. discoidalis it was 59.55 µg/L. In the oviposition of both species of argulids, maximum mortality began with treatments of 140 µg/L, while the LC50-24 hr for Argulus sp. and D. discoidalis was 83.98 µg/L and 82.48 µg/L, respectively. In both argulid species exposed to C. citratus essential oil, morphological alterations were observed only in the eyes, and they occurred in the ommatidium and rhabdomeres and were dependent on the concentration of C. citratus essential oil and the parasite species.

Additional morphological information on Dipteropeltis hirundo Calman, 1912, and a description of Dipteropeltis campanaformis n. sp. (Crustacea: Branchiura) from two characiform benthopelagic fish hosts from two Northern rivers of the Brazilian Amazon.

Dipteropeltis is a monotypic genus of the Branchiura and endemic to South America. Twelve specimens of Dipteropeltis sp. and micrographs of a thirteenth specimen were obtained from four institutions. Measurements and light micrographs were taken of all the specimens. The specimens were compared to all known descriptions of D. hirundo and sorted into two groups, those that conformed to the description of D. hirundo; and those that differed morphologically. Two specimens, one from each group¸ were stained with lignin pink and subsequently prepared for Scanning Electron Microscopy. The differences observed in the specimens indicated that a new species exists. Thus, Dipteropeltis campanaformis n. sp. is described from seven specimens collected from two characiform benthopelagic fish species and compared to D. hirundo. Dipteropeltis campanaformis n. sp. differs from D. hirundo in the head shape, the length and shape of the carapace lobes, the membrane composition of the maxillules, the shape of the maxillae, the shape of the mouth, and the shape of the natatory lobes.

A bridge between original and novel states: ontogeny and function of "suction discs" in the Branchiura (Crustacea).

The emergence of novel structures in the course of evolution faces an explanatory problem, leaving the gap from the ancestral structures difficult to bridge. This difficulty is caused by the lack of intermediate stages. Branchiurans are ectoparasitic crustaceans which use a pair of "suction discs" to attach to their host. These structures are modified first maxillae. During ontogeny, the first maxillae transform from a normal cephalic appendage to the specialized suction disc. However, supposedly ancestral branchiurans lack the suction discs in the adults and the first maxilla remains a normal appendage throughout. We describe the muscular arrangements in the developing first maxillae in Argulus coregoni. The suction discs originate as a fusion of the first and second podomeres. The sucker muscles of the suction discs are homologous to the muscles that insert in the second podomere at the early larval stages. The developmental process of the suction disc can be seen as a "recapitulation" of the evolutionary process. We thus show how the first maxilla can maintain not just the biological role but also a functional continuity during the evolution of the novel structure. From this example it is obvious that the intermediate stages of the emerging novelty, if present in the ontogeny, can help solve at least some of the enigmatic appearances of novel structures.

Argulus elongatus (Branchiura, Argulidae) in fish in the upper São Francisco river, Brazil.

Among 164 fish from the upper São Francisco river, caught in the Três Marias reservoir (18º 12' 59" S; 45º 17' 34" W) or downstream from this reservoir (18º 12' 32" S; 45º 15' 41" W) in 2007, 2008, 2016 and 2017, four specimens of Argulus elongatus Heller, 1857 were found, one specimen per fish, in the following host species: Brycon orthotaenia Günther (two fish parasitized out of 38 examined) and Salminus hilarii Valenciennes (one fish parasitized out of 45 examined) (both in Bryconidae); and Metynnis lippincottianus (Cope) (one fish parasitized out of 81 examined) (Serrasalmidae). This opportunistic ectoparasite deserves attention even in the case of low infestation in fish in a natural water system. This is the first record of a branchiuran species in B.orthotaenia; the host list of A.elongatus is extended through addition of the three fish species examined in this study, and the known Brazilian geographical distribution of this argulid is enlarged to the São Francisco river basin.

Dolops carvalhoi as a vector of Epistylis sp. between cultivated and wild specimens of Oreochromis niloticus in Brazil.

Epistyliasis has been characterized as an emergent disease which has a great impact on fish farms, especially on Oreochromis niloticus production systems. Although epidemiological important, information about the dispersion of these parasites and their mechanical vectors is scarce. The present study reported the cooccurrence of Epistylis sp. as an epibiont of Dolops carvalhoi, a parasitic crustacean of cultivated/wild specimens (from accidental release) of O. niloticus from a cage fish farm area in the Ilha Solteira Reservoir, Grande River, SP, Brazil. The co-occurrence of Epistylis sp. and D. carvalhoi, and their epibiont relationship registered in this study suppose that the Epistylis may use the crustaceans for dispersion and as mechanical vectors for the dissemination of diseases in wild and cultivated hosts. Moreover, exchange of parasites between wild and cultivated hosts is possible, considering both organisms (protozoan and argulid). Furthermore, the results of the present study demonstrate the need to monitor the areas adjacent to cage fish farms as a preventive measure for the dispersion of pathogens. This is the first report of epibiosis between Epistylis sp. and argulid parasites of cultivated and wild O. niloticus, contributing to knowledge about host-parasite specificity, geographical distribution, dispersion of etiological agents and epidemiology in aquaculture.

Histological and histochemical characterisation of glands associated with the feeding appendages of Argulus foliaceus (Linnaeus, 1758).

Argulus foliaceus (Linnaeus, 1758) is a member of the branchiuran family Argulidae, a group comprising parasitic "fish lice". A. foliaceus is distributed worldwide and causes major economic impacts for cultured freshwater fish globally. The work described in this study was undertaken with the objective of identifying, describing and characterising glands associated with feeding in A. foliaceus. From structural and ultrastructural microscopic studies of A. foliaceus, three types of gland were determined to be associated with the pre-oral spine and mouth tube and were suggested to be involved in feeding activities. Two of these glands, the labial glands and the proboscis glands, appeared to secrete their products via the mouth tube and a third, the spinal gland, was connected directly to the pre-oral spine. The current study confirmed that the pre-oral spine delivers active secretions from the spinal gland, which may aid in immunomodulation, while the tubular labial spines and proboscis glands openings within the mouth tube may serve to enhance the feeding process by delivering salivary components to aid pre-digestion and immune-modulate the host. The suggested functions are supported by histological and histochemical staining, coupled with fluorescent lectin-binding assays, which enabled characterisation of the carbohydrate moieties associated with these glandular tissues.

Chonopeltis australis (Crustacea: Branchiura); the female reproductive system.

The female reproductive system has been described for Dolops ranarum (Stuhlman, 1891) and various Argulus spp. but, there is no description of the reproductive system for Dipteropeltis spp. Calman, 1912 or Chonopeltis spp. Thiele, 1900. This paper describes the female reproductive system and egg laying behaviour of Chonopeltisaustralis Boxshall, 1976 using histology, light microscopy, scanning electron microscopy and observations. The histological study of six specimens showed that the organ structure is similar to that of Argulus spp. and D. ranarum. The oocytes therefore develop in the gonocoel, are ovulated into the lumen of the ovary which is continuous with the functional oviduct and eventually the gonopore. Females of C. australis deposit eggs on the surrounding substrate while the fish is at rest, without leaving the host, thereby mitigating the risk of not locating a host again.

Swimming and cleaning in the free-swimming phase of Argulus larvae (crustacea, branchiura)--appendage adaptation and functional morphology.

The free-swimming early larval stages of Argulus foliaceus (Linneaus) (Branchiura) are studied using digital video, light microscopy, and SEM. We analyze and document the mode of swimming in the hatching stage of A. foliaceus and the subsequent juvenile stages with fully developed thoracopods. We present new observations and an analysis of the functional morphology of a cleaning behavior in the first stage. This stage swims very efficiently using the large exopods of the second antennae in concert with the mandibular palp (naupliar limbs), while the subsequent stages use the now developed thoracopods for propulsion. This posterior shift in propulsion is similar to--but independent from--what is seen in other crustaceans. The hatching stage has previously been referred as a "metanauplius" but as the first and second maxillae are developed and active, and buds of all four thoracopods are present, it is too advanced to be included in the naupliar phase. The hooks of the first antennae and the distal hooks of the maxillae are demonstrated to function not only as attachment organs (to the host), but also to play a significant role in the cleaning of the naupliar swimming appendages. A digital video-based analysis of the swimming mode is provided. The larval swimming pattern is generally similar to that of other crustaceans such as Branchiopoda and Cirripedia, but autapomorphies of the Branchiura include the following: 1) While actively swimming, the naupliar appendages are almost straight during the recovery stroke and 2) they have a relatively small deflection during movement ( approximately 25 degrees or approximately 35 degrees for mandible and second antenna respectively), 3) the larval mandible has a uniramous palp which is the retained exopod. The morphological implications of the transition from the possibly nonfeeding pelagic, or free-swimming, first larval stage to the feeding, parasitic second stage are discussed and compared with other crustaceans.

Male reproductive system of the fish ectoparasite Argulus bengalensis (Crustacea: Branchiura).

The male reproductive system of the fish ectoparasite Argulus bengalensis was reconstructed with serial histological sections through longitudinal and transverse planes and the description was re-evaluated after the discovery of spermatophore. The study revealed that the testis of A. bengalensis consists of two lobes, each comprised the two enclaves. The outer enclaves from both lobes are connected proximally with an isthmus. The inner enclave is covered with two layers of chromatophores which are presumably involved in androgenic hormone production. At the proximal end of the inner enclaves a pair of vasa efferentia originates and ascends into thorax where it leads into vesicula seminalis. Like Dolops ranarum, a pair of spermatophoric glands is found in the thorax just behind the second maxilla. Glandular cells of the spermatophoric glands are columnar with a large nucleus at their basal side. A pair of spermatophoric canals carries the secretion of the glands into the vesicula seminalis through vasa efferentia. Spermatophores are delivered to the female through a median vas deference originating from the vesicula seminalis. In addition to the spermatophoric glands, a pair of accessory glands is located in the coxa of the fourth thoracic legs. At the region dorsal to the isthmus both the spermatophoric canal and accessory gland ducts unite with the vas efferens of the respective side. The secretion of the accessory gland presumably contributes to semen composition of the parasite.

Chonopeltis australis (Crustacea) male reproductive system morphology; sperm transfer and review of reproduction in Branchiura.

The morphology of the male reproductive system as well as sperm transfer in Branchiura has been described for Dolops ranarum and Argulus japonicus. In this study, the reproductive system and accessory structures are described for male Chonopeltis australis using histology, light microscopy, and scanning electron microscopy. For the first time, we describe sperm transfer by means of a spermatophore in this genus. The internal and external morphology and mechanism of sperm transfer is compared with other Branchiura, where it has been described. The morphology of the reproductive system of C. australis is similar to that of D. ranarum while the accessory structures and the spermatophore produced are similar to that of A. japonicus. A revision of the definition of Branchiura with respect to reproduction is provided.

Argulus elongatus (Branchiura, Argulidae) in fish in the upper São Francisco river, Brazil / Argulus elongatus (Branchiura, Argulidae) de peixes do alto rio São Francisco, Brasil

Abstract Among 164 fish from the upper São Francisco river, caught in the Três Marias reservoir (18º 12' 59" S; 45º 17' 34" W) or downstream from this reservoir (18º 12' 32" S; 45º 15' 41" W) in 2007, 2008, 2016 and 2017, four specimens of Argulus elongatus Heller, 1857 were found, one specimen per fish, in the following host species: Brycon orthotaenia Günther (two fish parasitized out of 38 examined) and Salminus hilarii Valenciennes (one fish parasitized out of 45 examined) (both in Bryconidae); and Metynnis lippincottianus (Cope) (one fish parasitized out of 81 examined) (Serrasalmidae). This opportunistic ectoparasite deserves attention even in the case of low infestation in fish in a natural water system. This is the first record of a branchiuran species in B.orthotaenia; the host list of A.elongatus is extended through addition of the three fish species examined in this study, and the known Brazilian geographical distribution of this argulid is enlarged to the São Francisco river basin.

Resumo De 164 peixes do alto rio São Francisco, capturados no Reservatório de Três Marias (18º 12' 59" S; 45º 17' 34" O) ou à jusante dele (18º 12' 32" S; 45º 15' 41" O), em 2007, 2008, 2016 e 2017, quatro espécimes de Argulus elongatus Heller, 1857 foram encontrados, sendo um branquiúro por peixe nas seguintes espécies de hospedeiros: Brycon orthotaenia Günther (dois peixes parasitados de 38 peixes examinados), Salminus hilarii Valenciennes (um peixe parasitado de 45 examinados) - ambos Bryconidae, e Metynnis lippincottianus (Cope) (um peixe parasitado de 81 examinados) - Serrasalmidae. Esse ectoparasito oportunista merece atenção, mesmo no caso de infestações baixas em peixes de ambientes hídricos naturais. Esse é o primeiro registro de Branchiura em B.orthotaenia. A lista de hospedeiros de A. elongatus é ampliada com a adição das três espécies de peixes examinadas neste estudo. A distribuição geográfica conhecida desse argulídeo no Brasil, é ampliada para a bacia do rio São Francisco.

Dolops carvalhoi as a vector of Epistylis sp. between cultivated and wild specimens of Oreochromis niloticus in Brazil / Dolops carvalhoi como vetor de Epistylis sp. entre espécimes cultivados e silvestres de Oreochromis niloticus no Brasil

Abstract Epistyliasis has been characterized as an emergent disease which has a great impact on fish farms, especially on Oreochromis niloticus production systems. Although epidemiological important, information about the dispersion of these parasites and their mechanical vectors is scarce. The present study reported the cooccurrence of Epistylis sp. as an epibiont of Dolops carvalhoi, a parasitic crustacean of cultivated/wild specimens (from accidental release) of O. niloticus from a cage fish farm area in the Ilha Solteira Reservoir, Grande River, SP, Brazil. The co-occurrence of Epistylis sp. and D. carvalhoi, and their epibiont relationship registered in this study suppose that the Epistylis may use the crustaceans for dispersion and as mechanical vectors for the dissemination of diseases in wild and cultivated hosts. Moreover, exchange of parasites between wild and cultivated hosts is possible, considering both organisms (protozoan and argulid). Furthermore, the results of the present study demonstrate the need to monitor the areas adjacent to cage fish farms as a preventive measure for the dispersion of pathogens. This is the first report of epibiosis between Epistylis sp. and argulid parasites of cultivated and wild O. niloticus, contributing to knowledge about host-parasite specificity, geographical distribution, dispersion of etiological agents and epidemiology in aquaculture.

Resumo Epistilíase tem sido caracterizada como uma doença emergente de grande impacto nas pisciculturas, especialmente em sistemas de produção de Oreochromis niloticus. Embora seja importante epidemiologicamente, informações sobre a dispersão desses parasitas e seus vetores mecânicos são escassos. O presente estudo relata a coocorrência de Epistylis sp. como um epibionte de Dolops carvalhoi, um crustáceo parasita de espécimes cultivados e silvestres (de escapes acidentais) de O. niloticus em área de piscicultura de tanques-rede no reservatório de Ilha Solteira, Rio Grande, SP, Brasil. A coocorrência de Epistylis sp. e D. carvalhoi, e sua relação epibionte registrada neste estudo, supõem que Epistylis sp. possa utilizar os crustáceos para dispersão e como vetores mecânicos para a disseminação de doenças em hospedeiros silvestres e cultivados. Além disso, a troca de parasitas entre hospedeiros silvestres e cultivados é possível, considerando ambos os organismos (protozoários e argulídeos). Os resultados deste estudo, também demonstram a necessidade de monitorar as áreas adjacentes às pisciculturas como medida preventiva para a dispersão de patógenos. Este é o primeiro relato de epibiose entre Epistylis sp. e parasitas argulídeos de O. niloticus cultivados e silvestres, contribuindo para o conhecimento da especificidade parasitária do hospedeiro, distribuição geográfica, dispersão de agentes etiológicos e epidemiologia na aquicultura.

Expanded description of Dolops bidentata (Bouvier, 1899) (Branchiura: Argulidae) based on specimens collected on Pygocentrus nattereri Kner, 1858 (Characiformes) from Poconé Wetland, MT, Brazil.

The current information on the branchiuran Dolops bidentata, a species described more one hundred years ago, is valid but incomplete; hence, an expanded description is given herein. Additional morphological information was obtained by light and scanning electron microscopy from specimens collected on Pygocentrus nattereri from the Poconé Wetland, MT, Brazil. Description of the appendages and other structures such as respiratory area, mouth, details and ornamentation of antennules and maxillae are provided for the first time.

The ultrastructure of the digestive cells of Argulus japonicus, Thiele 1900 (Crustacea: Branchiura).

The ultrastructure of the cells of the digestive system of Argulus japonicus is described with the use of transmission electron microscopy. Specimens of Argulus japonicus were collected from the Vaal Dam in South Africa and fixed in Todd's fixative. The samples were post fixed in osmium tetroxide and embedded in resin. The anterior midgut is composed mostly of R cells while the enteral diverticula are composed mainly of R cells in the proximal diverticules and of F cells in the distal diverticula. The posterior midgut is composed of very large papilliform B cells and of R cells. The R cells in the anterior midgut probably absorb nutrients including lipids. The F cells are filled mostly with rough endoplasmic reticulum, suggesting enzyme synthesis, while the B cells portrayed endocytotic vesicles, indicating intracellular digestion of predigested food. The R cells of the posterior midgut are less active than cells present in the anterior midgut. E cells and peritrophic membrane were not observed.

The effect of starvation on the ultrastructure of the digestive cells of Dolops ranarum (Stuhlmann, 1891) (Crustacea: Branchiura).

Transmission electron microscopy was conducted on the digestive epithelium of the crustacean ectoparasite Dolops ranarum to elucidate its ultrastructure for the first time, both in a nourished and starved condition. Specimens were collected from the Limpopo Drainage System in South Africa, and the specimens were killed and dissected in Todd's fixative. The anterior midgut is composed mostly of absorptive cells or R cells, while the diverticula are composed of R cells and of F cells, which are moderately abundant in rough endoplasmic reticulum. They are probably responsible for producing digestive enzymes. The posterior midgut is composed of papilliform B cells with large apically located vesicles and R' cells devoid of cell inclusions. Under starvation, specimens survive for a maximum of 12 days; R cells show the most conspicuous changes in ultrastructural characteristics. It is concluded that D. ranarum has adapted to short-term survival only without a host.

Ultrastructure of Argulus japonicus Thiele, 1900 (Crustacea: Branchiura) collected from Guangdong, China.

Argulus japonicus Thiele, 1900 was originally described from China, but has spread to Europe, Africa, Australia, and North America. An ultrastructural description of A. japanicus has been done as one of the most common fish ectoparasites which has a high affect on fish life. The description was based on light and scanning electron microscopes studies using 13 females and 7 males which were collected from Shunde city in Guangdong Province, China. Typical Argulus body and body's appendages are associated with our specimens, carapace covers fourth leg, and scales separate uniformly on frontal and lateral lobes, ventral surface of thorax is ornamented by scales contrary of the dorsal surface which is quite smooth, various shapes of scales have been detected on second maxillae. Location and structure of copulatory accessory, number and structure of overlapping sclerites of sucking disk rim, melanophores patches of male, second swimming leg structure, and parasite's total size are the most important criteria which may be used to distinguish between male and female. Moreover, new classification criteria of Argulide, obtained by scanning electron microscopic study of previous species, are discussed.

Redescription of Argulus multipocula Barnard, 1955 (Crustacea: Branchiura) collected on the west coast of South Africa.

Argulus multipocula Barnard, 1955 was originally described from a single female found in a littoral sample from the east coast of South Africa. We present a redescription of this species based on light and scanning electron microscope studies of 25 females collected from the southern mullet Liza richardsonii (Smith) on the west coast of South Africa.