Structure of the male reproductive accessory glands of Pterostichus nigrita (Coleoptera: Carabidae), their role in spermatophore formation

Accessory gland secretions of male insects have many important functions including the formation of spermatophores. We used light and electron microscopy to investigate the structure of the accessory glands and posterior vasa deferentia of the carabid beetle Pterostichus nigrita to try to determine where spermatophore material is produced. Each accessory gland and posterior vas deferens had an outer layer of longitudinal muscle, beneath which was a layer of connective tissue and a thin band of circular muscle, all of which surrounded a layer of epithelial cells lining the lumen of the ducts. Based on the ultrastructure of the epithelial cells, and their secretory products, we identified two epithelial cell types in each region (distal and proximal) of the accessory glands and four types in the posterior vas deferens. Most secretory products, which stained positively for proteins and some mucins, were released into the lumen of the ducts by apocrine secretion. The accessory glands produced one type of secretory product whereas in posterior vasa deferentia, four types of secretory products were found layered in the lumen. Our results suggest that most of the structural material used to construct a spermatophore is produced by the cells of the posterior vasa deferentia.     

Histochemical characteristics of spermatophore layers of Scylla serrata (Forskal) (Decapoda: Portunidae)

The spermatophores of S. serrata are protected by an outer thick chitinous layer and an inner thin non-chitinous one. Both layers are rich in acid mucopolysaccharides containing sulphated (outer layer) and carboxylic groups (inner layer). The proteins of the two layers show much tryptophan, but lack tyrosyl, sulfhydryl and disulphide groups. No phenols or phenol oxidases could be detected histochemically in either layer, suggesting the absence of phenolic tanning in the spermatophore. The physical properties, as revealed from treatment with acids and alkali, indicate the resistant nature of the outer layer; the inner layer easily shrinks or disrupts under such treatment. The outer layer, though resistant, is readily permeable to low molecular weight dye substances employed in permeability experiments. The mechanism of sperm release is recorded and discussed. It is suggested that, in S. serrata, the dehiscence of spermatophore may be caused by imbibing of low molecular weight substances by the sperm mass substances of the spermatophore while the latter is inside the spermatheca.     

Observations on the ixodid ticks associated with domestic ungulate animals in Jordan

Abstract

Fourteen species and subspecies of ixodid ticks, representing four genera, were collected from native sheep, goats, camels and cattle in Jordan. Haemaphysalis otophila was the most common species (48.1%) with Rhipicephalus sanguineus (15.3%), Boophilus kohlsi (14.4%) and Haemaphysalis sulcatus (11.6%) representing other common species among the 9,545 specimens examined.     

A method for extraction and analysis of high quality genomic DNA from ixodid ticks

Currently, no published methods describe the extraction of high molecular weight genomic DNA from ixodid ticks (Acari: Ixodidae) and commonly used methods of extraction are not well adapted for use with members of this family. A method for extraction of minimally degraded genomic DNA from ixodid ticks that can be completed in one or two days is described. The method produces DNA which is of sufficient size (>24 kb) for use in Southern analysis and which is readily digestible by restriction endonucleases. Southern analysis using a cytochrome P450 gene probe, demonstrates the success of our method with genomic DNA extracted from two species of Ixodidae, the lone star tick, Amblyomma americanum (Linnaeus) and the cattle fever tick, Boophilus microplus (Canestrini).     

New insights in the male anatomy,spermatophore formation,and sperm structure in Atyidae: The red cherry shrimp Neocaridina davidi

This study aims to analyze the functional anatomy of the male reproductive system in Neocaridina davidi, a very popular ornamental species of caridean shrimp. Mature males were cold‐anaesthetized and their reproductive systems were dissected for histological and histochemical analysis, while the spermatozoa and spermatophore wall ultrastructure were analyzed under transmission electron microscopy. The male reproductive system consisted of two coiled testes, which were continuous with the vasa deferentia. Testes were positioned on the dorsal side of the cephalothorax above the hepatopancreas, and comprised seminiferous tubules where spermatogenesis occurred. Each vas deferens (VD) was a long tube dorsolaterally positioned with respect to the hepatopancreas, and increased in diameter at the distal end. Three regions could be recognized in the VD: proximal, middle, and distal. The proximal region had a cylindrical epithelium with secretory cells. The middle region (or typhlosole) had a dorsal fold (or typhlosole) with a thick columnar epithelium and high secretory activity. The spermatophore was a continuous cord with three acellular layers, which were mainly characterized by the presence of neutral glycoconjugates and proteins. The sperm morphology was distinct from the inverted cup‐shaped spermatozoa observed in the majority of caridean shrimps. The spermatozoa in specimens of N. davidi were spherical in shape, with a cross‐striated, single, short spike, and arranged in clusters of three or four sperm cells. The composition of the spermatophore, and the arrangement and form of the spermatozoa, seem to be unique in comparison to other species of Caridea.     

Spermatophoric reaction reappraised: Novel insights into the functioning of the loliginid spermatophore based on Doryteuthis plei (Mollusca: Cephalopoda)

During copulation, spermatophores produced by male coleoid cephalopods undergo the spermatophoric reaction, a complex process of evagination that culminates in the attachment of the spermatangium (everted spermatophore containing the sperm mass) on the female's body. To better understand this complicated phenomenon, the present study investigated the functional morphology of the spermatophore of the squid Doryteuthis plei applying in vitro analysis of the reaction, as well as light and electron microscopy investigation of spermatangia obtained either in vitro, or naturally attached on females. Hitherto unnoticed functional features of the loliginid spermatophore require a reappraisal of some important processes involved in the spermatophoric reaction. The most striking findings concern the attachment mechanism, which is not carried out solely by cement adhesive material, as previously believed, but rather by an autonomous, complex process performed by multiple structures during the spermatophoric reaction. During evagination, the ejaculatory apparatus provides anchorage on the targeted tissue, presumably due to the minute stellate particles present in the exposed spiral filament. Consequently, the ejaculatory apparatus maintains the attachment of the tip of the evaginating spermatophore until the cement body is extruded. Subsequently, the cement body passes through a complex structural rearrangement, which leads to the injection of both its viscid contents and pointed oral region onto the targeted tissue. The inner membrane at the oral region of the cement body contains numerous stellate particles attached at its inner side; eversion of this membrane exposes these sharp structures, which presumably adhere to the tissue and augment attachment. Several naturally attached spermatangia were found with their bases implanted at the deposition sites, and the possible mechanisms of perforation are discussed based on present evidence. The function of the complex squid spermatophore and its spermatophoric reaction is revisited in light of these findings. J. Morphol. 2012. © 2011 Wiley Periodicals, Inc.     

Prevalence of ixodid ticks on cattle in Mazandaran province, Iran

A survey was carried out to investigate the prevalence of hard tick species (Acari: Ixodidae) on cattle in Mazandaran province, Iran. A total of 953 ticks were collected from 86 infested cattle during activating seasons of ticks during 2004-2005. Nine species were identified: Boophilus annulatus (51.3%), Rhipicephalus bursa (16.8%), Haemaphysalis punctata (6.3%), Ixodes ricinus (6.8%), Hyalomma marginatum (12.5%), Hyalomma anatolicum excavatum (5.2%), Hyalomma asiaticum (0.6%), Hyalomma detritum (0.2 %), and Dermacentor spp. (0.1%). The results show that Boophilus annulatus, Rhipicephalus bursa, and Hyalomma species are dominant tick species in the surveyed area.     

Mating effort function of the spermatophore in the bushcricket Poecilimon veluchianus (Orthoptera, Phanero[pteridae): support from a comparison of the mating behaviour of two subspecies.

To analyse spcrmatophore function, various aspects of the mating behaviour (e.g. spermatophore mass, duration of sperm transfer, mating frequency) were compared in two subspecies of the bushcricket P. veluchianus. Body mass was significantly different in both subspecies and had a strong effect on spermatophore mass, resulting in a large difference in absolute and relative (percentage of male body mass) spermatophore mass in both subspecies. After copulation the small P. v. minor spermatophores were consumed much faster by the female than the larger ones of P. v. veluchianus. The time necessary for sperm transfer from the spermatophore to the female spermatheca was much shorter in the subspecies with small spermatophores than in that with large spermatophores, and in both subspecies similar to the time required to consume the spermatophores. Mating frequency varied also between the subspecies and was murh higher in P. v. minor than in P. v. veluchianus. Differences in body mass between the two subspecies therefore resulted in changes in several aspects of mating behaviour. However, despite large differences in the mass of the spermatophore, its main function seems to be to ensure sperm transfer. This can be concluded from the similarity between the duration of sperm transfer and the time period necessary for spermatophore consumption in each of the subspecies. The spermatophore is thus considered to be male mating effort.     

Hard ticks (Acari,Ixodidae) of Croatia

The present paper is based on original and literature data. In Croatia the first studies on the occurrence of ixodid species were made about 80 years ago. The number of tick species recorded in Croatia considerably increased during the 1950s, 60s, 70s and 80s of the past century. A total of 21 species of hard tick belonging to 5 genera have been recorded in Croatia. Ixodes is the best represented genus, with seven species recorded. Haemaphysalis is represented by six species, followed by Rhipicephalus with four species. Dermacentor and Hyalomma are represented by two species each. The ticks were collected on 47 different host species. Eleven tick species were collected on Bos taurus and Ovis aries, followed by Capra hircus and Equus caballus with 8 species and Canis lupus familiaris with 6 species. On the remaining 42 host species one, two or three tick species were collected. The most widespread tick is Ixodes ricinus which was found on 25 different host species.     

Long hyaline gland discharge and multiple spermatophore formation by the male grasshopper, Melanoplus sanguinipes

ABSTRACT. Discharge from the male accessory reproductive gland (ARG) by the male grasshopper, Melanoplus sanguinipes (Fabr.), has been studied by assay of a characterized product, a glycoprotein LHPI, and the rate of formation of spermatophores. LHPI is an exclusive long hyaline gland (a prominent ARG tubule type) product whose discharge is symmetrical from the bilaterally paired glands. LHPI forms 65% of a viscous secretion that is discharged concomitantly with the spermatophores. Though low ARG reserves in 5-day-old males limit both the number of spermatophores formed and LHPI discharged in copulations ≤6 h, this appears to be the result of shorter copulations. The number of spermarophores formed in 1 h was not impaired by general depletion of ARG protein (by repeated copulations) or by selective depletion of long hyaline gland protein (by unilateral and bilateral long hyaline gland removal), though these manipulations reduced LHPI discharge by 22%, 44% and 100%, respectively. However, 56% of spermatophores formed by males with the long hyaline gland bilaterally ablated failed to uncoil properly. These results indicate LHPI and/or other long hyaline gland proteins may act as lubricants. Unlike spermatophore formation and LHPI discharge, which increased steadily up to 90–120 min then levelled off, transfer of radiolabelled male ejaculate to the spermatheca was very variable. In 90 min copulations, only 1% of the total radioactivity (representing c. 5 μg protein) lost from the ARG complex was transferred to the spermatheca. The importance of male-derived protein in vitellogenesis is discussed.     

Development of the digestive system of Argentine hake,Merluccius hubbsi,larvae

The Argentine hake Merluccius hubbsi is an important fishery resource of the Southwestern Atlantic Ocean and it is also a potential species for cultivation. In this work, the digestive system development in field-caught hake larvae was studied using histological and histochemical approaches. The digestive tract of larvae was divided into: oropharyngeal cavity (OPC), esophagus, stomach (that develops in the preflexion stage), and intestine. The annexed digestive glands consisted of the liver and the exocrine pancreas. At the beginning of the preflexion stage, teeth were developed in the OPC. There were mucous cells in the esophagus secreting different glycoconjugates from hatching. The enterocytes in the posterior intestine exhibited supranuclear vesicles associated with protein absorption. Mucous cells were observed in the posterior intestine in the preflexion stage and, in the anterior region, ending the flexion stage. Each type of glycoconjugates has a specific role. Acidic mucins lubricate and protect from mechanical damage, sialomucines protect from bacterial infections and neutral mucins regulate the acidity of mucus secretion, protect against abrasion and participate in the formation of the chyme, indicating a pregastric digestion. The liver was present since hatching with pancreatic tissue inside and increased in size acquiring the typical structure with hepatocyte cords, sinusoids, vacuoles, and hepatic duct. The hepatocytes vacuolization increased with larval development. The pancreas became extra-hepatic, with basophilic acinar cells and acidophilic zymogen granules. Throughout the ontogeny, the increased structural and functional complexity of the digestive system reflected the transition to exogenous feeding and nutritional increasing needs.     

Structural changes in the spermatophore of the freshwater 'red claw' crayfish Cherax quadricarinatus (Von Martens, 1898) (Decapoda, Parastacidae)

López Greco, L.S. and Lo Nostro, F.L. 2007. Structural changes in the spermatophore of the freshwater ‘red claw’ crayfish Cherax quadricarinatus (Von Martens, 1898) (Decapoda, Parastacidae). —Acta Zoologica (Stockholm) 88 : 000–000 The structure of the spermatophore was studied in Cherax quadricarinatus. Pieces of the distal vas deferens and transferred spermatophore from the females were fixed, cut and stained. Within the distal vas deferens, the primary layer and the secondary layer of the spermatophore were distinguishable. In the latter, two components were detected: cytoplasmic droplets and a homogeneous matrix. During the first 10 minutes post‐extrusion the cytoplasmic droplets drastically changed from looking like ‘empty droplets’; at this time the spermatophore changed from a liquid stage to a sticky one. One hour after extrusion the spermatophore began to harden and within the first 24–48 h post‐mating it was a solid and intense white structure tightly attached to the female; after 72 h it acquired a softer aspect, completely dehiscing between 96 and 120 h post‐mating. Histologically, the primary layer maintained its integrity surrounding the spermatozoa while the secondary layer lost the cytoplasmic droplets. The spermatophore began to hydrate between 24 and 48 h and by 72–96 h many sections of the sperm cord began to coalesce. From 48 h post‐mating some fissures appeared within the matrix that enlarged between 72 and 120 h. We propose that both manipulation by the female and hydration are the mechanisms involved in the release of the spermatozoa from the spermatophore.     

Protein analysis of the spermatophore reveals diverse compositions in both the ampulla and the spermatophylax in a bushcricket

Nuptial gifts are male mating investments, which, in bushcrickets, comprise the sperm‐containing ampulla and the attached spermatophylax. The functions of the spermatophylax are to deter females from premature removal of the sperm‐containing ampulla, which is a nutrient resource for females, as well as a source of compounds that influence female behaviour to increase male evolutionary fitness. Placing these functions into a proteomic perspective, we analyze the protein composition of nuptial gifts from male Poecilimon ampliatus (Brunner von Wattenwyl , 1878) bushcrickets using large two‐dimensional gel electrophoresis coupled with nano‐liquid chromatography‐electrospray ionization mass spectrometry and de novo sequencing. We separate the proteins with high resolution and detect approximately 600 protein spots in the seminal fluid (ampulla) and 300 in the spermatophylax. There is only a small fraction of overlap in protein spots, whereas the majority differ between the two compartments. As a result of the lack of a sequenced genome and protein data for this non‐model insect, we are unable to identify the proteins. We discuss the diversity of proteins, as well as their size range, in light of potential protein costs and potential functions.     

Distribution and phenology of ixodid ticks in southern Zambia

Distribution data for epidemiologically important ticks (Acari: Ixodidae) in the Southern Province of Zambia, one of the main cattle areas of the country, are presented. Boophilus microplus (Canestrini) was not recorded in southern Zambia, whereas Boophilus decoloratus (Koch) is present throughout the area. New distribution patterns for less economically important ixodid ticks are also discussed. Southern Zambia is a transition zone because it is the most northern area in Africa where mixed Rhipicephalus appendiculatus Neumann and Rhipicephalus zambeziensis Walker, Norval & Corwin populations were reported. Although a second generation of adult R. appendiculatus/R. zamnbeziensis was encountered, simulations indicated that this phenomenon is very rare in southern Zambia, mainly because of the colder temperatures during the early dry season and lower rainfall. These simulations were supported by a development trial under experimental conditions. Tick body size measurements showed that southern Zambian ticks are larger than eastern Zambian R. appendiculatus. It is hypothesized that body size is related to diapausing intensity in this species. The epidemiological consequences are that a different approach to control Theileria parva (Theiler) (Piroplasmida: Theileriidae) and other tick-borne diseases is needed in southern Zambia, compared to the one adopted in eastern Zambia.     

Risk factors associated with ixodid tick species distributions in the Basque region in Spain

Ixodid tick abundance was investigated in the Basque region in Spain in two 1-year longitudinal studies, in 1992-1993 and 2003-2004. Forty zones were visited monthly and 162 672 ticks (87% larvae, 12% nymphs and 1% adults) were collected by blanket dragging. Eleven tick species belonging to the genera Ixodes, Haemaphysalis, Rhipicephalus and Dermacentor were identified including Haemaphysalis concinna Koch, which had not previously been reported in Spain. Tick species abundance differed between zones, studies and seasons. In 1992-1993, Haemaphysalis punctata Canestrini & Fanzago was the predominant species and distinct spring-summer and autumn-early winter peaks of activity were observed. In 2003-2004, Ixodes ricinus (Linneaus) was the most common species and was active throughout the winter. Larval and nymph seasonal activity patterns coincided in both 1993 and 2003 and this could facilitate co-feeding transmission of pathogens. Higher tick abundance was associated with increased livestock abundance in 1992-1993 and milder winter temperatures in 2003-2004. Tick collection rates in areas with moderate and high tick density were positively associated with the interaction between ambient temperature at sampling and rainfall 7 days prior to sampling. Collection rates were also significantly higher at medium rather than higher altitude, in forested areas than in open grasslands and lower in recreational areas frequented by people and with wet vegetation at sampling.     

A review of the types of parasite-host relations in ixodid ticks

In the first type of mutual relationships the ixodid ticks expose an evident pathogenicity to vertebrate hosts (tick toxicoses) and in the second type the hosts expose a resistance on a base of organism immune system activisation in a response to blood sucking. In the third type of relationships an interaction between the ticks and hosts have a dynamic character (tolerability of host). The first type of relationships leading to a selective elimination of hosts according to our opinion means a comparatively youth of relationships. The third type represent ancient and long term relationships. The second type probably having been widely spread in a historical past occupies an intermediate position. Depending upon areal the ticks of the same type expose different degree of the pathogenicity. Even an acquired resistance of the host does exist in natural conditions, it is a transient phenomenon.     

Shifts in the distribution of ixodid ticks parasitizing cattle in Zimbabwe

In an attempt to update information on the ecological distribution of ixodid ticks (Ixodida: Ixodidae) in Zimbabwe, a cross‐sectional survey was carried out between September 2013 and May 2015 at 322 dip tanks. A total of 15 tick species were collected, namely: Amblyomma hebraeum Koch (65.2%, n = 210/322), Amblyomma variegatum Fabricius (14.9%, n = 48/322), Hyalomma rufipes Koch (62.4%, n = 201/322), Hyalomma truncatum Koch (37.9%, n = 122/322), Rhipicephalus appendiculatus Neumann (60.6%, n = 195/322), Rhipicephalus compositus Neumann (0.3%, n = 1/322,), Rhipicephalus decoloratus Koch (61.8%, n = 199/322), Rhipicephalus evertsi evertsi Neumann (65.2%, n = 210/322), Rhipicephalus lunulatus Neumann (4%, n = 13/322), Rhipicephalus microplus Canestrini (32%, n = 103/322), Rhipicephalus near punctatus Walker and Horak (7.1%, n = 23/322), Rhipicephalus simus Koch (5.6%, n = 18/322) and Rhipicephalus cf. turanicus Pomerantsev (3.4%, n = 11/322). Compared with previous surveys, changes in the distribution of A. hebraeum, A. variegatum and R. microplus were recorded. The distributions of other tick species have largely remained unchanged. Factors which might have influenced these changes and the possible impacts on the epidemiology of tick‐borne diseases are discussed.