Maud Bonato

Investment in eggs is influenced by male coloration in the ostrich, Struthio camelus

Life history theory predicts that females should modify their investment in a particular breeding attempt according to the likelihood of its success, as the investment of females in reproduction is typically higher than that of males. The ostrich mating system is promiscuous, and is thus a particularly interesting one in which to investigate differential investment by the sexes. To date, there has been no evidence that female ostriches discriminate between males as potential mates, but the degree of dimorphism in this promiscuous species and the variation in chick size within clutches suggest that differential maternal investment is likely. We investigated the relationship between egg mass and coloration of the feathers, bill, neck and legs of 15 male ostriches, maintained in a breeding flock at an ostrich farm in South Africa. Paternity was determined using microsatellite markers. We found that the colour of the neck, white and black body feathers, and the brightness of black feathers, predicted egg mass. These traits are exposed during the male courtship display, so we suggest that these visual cues influence the degree of maternal investment in eggs through their influence on female perception of mate quality.

Male coloration reveals different components of immunocompetence in ostriches, Struthio camelus

It has been suggested that secondary sexual ornamentation signals male ability to resist infections, as only high-quality individuals are able to invest both in high immune defence and elaborate ornament expression. Such ornaments could thus serve as indicators of male quality and could be used by females in choosing mates. Ostriches are sexually dimorphic with regard to coloration of their feathers, bill, neck and legs, and have a promiscuous mating system, with a high degree of reproductive skew, particularly in males. We investigated the relationship between the coloration of the feathers, bill, neck and legs of 15 male ostriches, maintained in a breeding flock, and the cell-mediated (measured using a phytohaemagglutinin (PHA) injection) and humoral components of their immune systems, as well as their heterophil:lymphocyte ratio. We found that male responses to PHA injection and humoral responses to tetanus were predicted by leg coloration, humoral responses to diphtheria were predicted by white feather coloration, and the heterophil:lymphocyte ratio was related to bill coloration. These traits, which relate to male immune capacity, are exposed during male–male interactions and courtship display, so we suggest that these visual cues could provide valuable information on male quality to females (as well as rival males), forming the basis of mate choice in this species.

Twice daily collection yields greater semen output and does not affect male libido in the ostrich

The success of an artificial insemination program in ostriches is highly dependent on the yield of viable semen. We, therefore, tested how semen output is affected by three different collection frequencies: once every 2d (48h interval), daily (24h interval), and twice a day (6h interval). Ejaculates were collected from seven male ostriches (aged 2-4 years) for 10 consecutive days using the dummy female method. We assessed semen characteristics (sperm motility, volume, concentration, number of sperm per ejaculate and sperm viability) and male libido (the delay between the presentation of the dummy and ejaculation, and the willingness to mount the dummy). The total daily output of semen and the number of sperm were greater at the 6h collection interval than at the 24h or 48h interval while sperm motility and viability were not affected. At the 6h interval, the number of live normal sperm increased over the treatment period while the number of live abnormal sperm was reduced. Furthermore, the time that males took to mount the dummy and their willingness to copulate with the dummy were unaffected by collection frequency. Across males we observed great individual variation in both semen characteristics and libido suggesting there is the potential to increase the efficiency of semen collection by selecting superior males. These results indicate not only that two collections per day yield maximum semen output and may improve semen viability, but also that quantifying variation between males may help further increase semen collection efficiency.

The development of ratite production through continued research

This paper summarises research on farmed ratites and their industries over the past 100 years. Commercial ratite products include meat, skins, feathers and oil. Research on ratites has attempted to enhance the quantity and quality of these products by focusing on the disciplines of breeding and genetics, reproduction and incubation, assisted reproduction, nutrition and animal welfare. Advances in these disciplines are discussed, and directions for future research are provided.

Ostrich chick humoral immune responses and growth rate are predicted by parental immune responses and paternal colouration

One of the most important measures of offspring performance is growth rate, which is often traded off against another important survival trait, immune function. A particular feature of ostrich chicks maintained in farmed environments is that cohorts of chicks vary widely in size. As parents can have a profound effect on the phenotype and fitness of their offspring, we investigated whether chick growth and immune defence were related to variation in levels of immune defence in their genetic parents. As secondary sexual traits of sires could serve as indicators of male quality, and be used in female mating decisions, we also investigated whether chick growth rate and immune defence were related to male plumage and integumentary colouration. We found that offspring growth rates and humoral responses were related to the humoral responses of their parents, suggesting that at least some components of humoral immune capacity are heritable. The white colour of male ostrich feathers was correlated to the humoral response and growth rate of their offspring, suggesting that this visual cue involved in the male courtship display could serve as an important signal to females of male quality, thereby forming the basis of mate choice in this species.

The effect of temperature and pH on the motility and viability of ostrich sperm

As the chemical environment of semen diluents can have a profound effect on sperm quality, we examined the effect of temperature and pH on the motility and viability of sperm in the ostrich. Semen was collected from four males, each male being replicated three times. Ejaculates were diluted and incubated for 10 min at 20°C and 40°C in four different buffers, temperature adjusted at pH 6, 7, 8 and 9 respectively. Average path velocity (VAP), curvilinear velocity (VCL), straight-line velocity (VSL), linearity (LIN), beat cross frequency (BCF) and amplitude of lateral displacement (ALH) were then recorded for each sample using CASA. The viability of sperm was assessed using nigrosin-eosin staining. Sperm incubated at 40°C had higher motility parameters, except for ALH. At 40°C, VAP, VSL and LIN increased with pH while VCL, BCF and ALH were higher for lower pHs. The viability of sperm was not affected by temperature but decreased at pH values>7. A pH in the neutral range appeared to yield higher quality sperm after in vitro storage at 20°C. However, the effect of different pH levels and temperatures on sperm longevity needs to be investigated further to develop viable ostrich specific diluents.

Predicting ejaculate quality and libido in male ostriches: Effect of season and age.

The success of artificial breeding program depends largely on the reproductive performance of males. Male performance can vary with season and age impacting on quality and quantity of semen collected for artificial insemination purposes and therefore fertility of inseminated females. We examined variation in semen output and male libido of seven male ostriches (aged 2-5 years) over a period of 24 months. We collected ejaculates using a dummy female and measured semen characteristics (ejaculate volume, sperm concentration, number of spermatozoa per ejaculate, sperm motility and morphology) and male libido (willingness to mount the dummy). A total of 1006 ejaculates were collected. Across months, the volume of semen (mean ± SEM) ranged from 1.03 ± 0.12 mL to 1.85 ± 0.07 mL, the sperm concentration from 3.21 ± 0.12 × 10(9)/mL to 4.16 ± 0.74 × 10(9)/mL, and the number of spermatozoa from 3.42 ± 0.28 × 10(9) to 7.66 ± 0.47 × 10(9). The largest volume of ejaculates and the highest number of sperm were collected in spring. Ejaculates with higher number of normal sperm were also collected in spring-early summer, whereas ejaculates with higher numbers of live abnormal and dead sperm were collected in winter. Sperm motility was relatively constant over months, despite a reduction in summer (January-February), while male libido peaked in winter (June-July) and spring (October-November). Furthermore, we observed high individual variation between males for all variables tested, except for motility. These results indicate that collections conducted in spring yield higher number of spermatozoa, when the libido of males is also at a maximum. Therefore in this species seasonal variation in semen quality should be considered in breeding programmes by artificial insemination to maximise fertility.

Classification of ostrich sperm characteristics

The success of assisted reproduction techniques is dependent on a sound foundation of understanding sperm characteristics to evaluate so as to improve semen processing. This study offers a descriptive basis for ostrich semen quality in terms of sperm function characteristics (SFC) that include motility, measured by computer assisted sperm analysis CASA (SCA(®)), viability (SYBR14/PI) and membrane integrity (hypo-osmotic swelling test). Relationships among these SFCs were explored and described by correlations and regressions. Certain fixed effects including the dilution of semen, season, year and male associated with semen collection were interpreted for future applications. The seasonal effect on sperm samples collected throughout the year suggested that it is prudent to restrict collections to spring and summer when SFCs and sperm concentration are maximized, compared to winter when these aspects of sperm quality are suppressed. Dilution of ejaculates helped to maintain important SFCs associated with fertilization success. The SFCs and sperm concentration varied among males, with specific males, having greater values for the percentage of motile (MOT) and progressively motile (PMOT) sperm, as well as sperm velocity (VCL, VSL, VAP) and linearity (LIN) variables. Males may thus be screened on these variables for inclusion in an artificial insemination (AI) programme to optimize fertility success rates.

A morphological classification of sperm defects in the ostrich (Struthio camelus)

Commercial ostrich farming is constrained by the absence of a formal animal recording and evaluation scheme as well as by current farming practices. Artificial insemination may have an important role in overcoming these limitations, but requires a thorough knowledge of sperm morphology. Although the morphological characteristics of normal ostrich sperm have been documented, little information is available on the incidence and structural peculiarities of defective sperm in this species. Semen smears were prepared from the ejaculates of five ostriches (Struthio camelus), stained and evaluated. Defects were observed in 17% of sperm studied. Tail defects constituted the most common anomaly. Various forms of bending were the main tail defect, ranging from gentle to acute bends of the principal piece, Dag-like coiling at the head base, sharp reflexes of the midpiece as well as coiling of the endpiece. In contrast, head defects were comparatively low in frequency, with macrocephalic sperm being the defect most frequently observed in this region. Bent, microcephalic, acephalic and round sperm heads were also noted but were few in number. Cytoplasmic droplets occurred frequently in the fixed smears, either associated with the sperm or as free-lying droplets. A small percentage of sperm with multiple defects was recorded. The incidence of morphologically normal sperm in ostrich semen compared favorably with that reported in emu semen, another commercially farmed ratite. However, the range of defects differed appreciably between the two species. Sperm tail anomalies were the most frequent category in the ostrich, whereas head defects comprised the main grouping in the emu.

Ostrich specific semen diluent and sperm motility characteristics during in vitro storage.

The dilution of semen is a very important initial process for semen processing and evaluation, storage and preservation in vitro and efficient artificial insemination. The aim of the study was to evaluate the effect of two synthetic diluents (OS1 and OS2) on ostrich sperm motility parameters during in vitro storage. Formulation of OS1 was based on macro minerals (Na, K, P, Ca, Mg) and OS2 on the further addition of micro minerals (Se and Zn), based on mineral concentration determined in the ostrich seminal plasma (SP). Sperm motility was evaluated at different processing stages (neat, after dilution, during storage and after storage) by measuring several sperm motility variables using the Sperm Class Analyzer® (SCA). Processing (dilution, cooling and storage) of semen for in vitro storage purposes decreased the values for all sperm motility variables measured. The percentage motile (MOT) and progressive motile (PMOT) sperm decreased 20% to 30% during 24 h of storage, independent of diluent type. Quality of sperm swim (LIN, STR and WOB), however, was sustained during the longer storage periods (48 h) with the OS2 diluent modified with Se and Zn additions. Quality of sperm swim with use of OS1 was 6% to 8% less for the LIN, STR, and WOB variables. Male fitted as a fixed effect accounted for >60% of the variation for certain sperm motility variables (PMOT, MOT, VCL, VSL, VAP and ALH) evaluated at different processing stages. Semen from specific males had sustained sperm motility characteristics to a greater extent than that of other males during the 24-h storage period.